Figure 1.
Shows that preincubation of immature brain cortical neurons with α-tocopherol (α-T) at nanomolar concentrations for 0.5 h prior to exposure of the cells to 0.2 mM H2O2 for 24 h did not increase the viability of brain cortical neurons (A); while preincubation with 10 and 100 nM α-T for 18 h prior to exposure of the neurons to 0.2 mM H2O2 for 24 h caused a pronounced increase in cortical neuron viability (B). H2O2 is designated as HP in the figure. Lactate dehydrogenase (LDH) method was used to determine neuron viability. The data bars (from the left to the right) in (A,B) show: (1) control values of LDH release from neurons; (2) control values of LDH release after incubation of the neurons with 100 µM α-T for 30 min (A) or for 18 h (B); (3) LDH release after neuron exposure to H2O2; (4) LDH release after preincubation of the neurons with 100 µM α-T prior to the cell exposure to H2O2; (5) LDH release after preincubation of the neurons with 100 nM α-T prior to the cell exposure to H2O2; (6) LDH release after preincubation of the neurons with 10 nM α-T prior to the cell exposure to H2O2. The results of one typical experiment from 9 experiments performed are shown as means ± SEM from 2–3 determinations in parallel samples. One way ANOVA followed by Tukey’s multiple comparison test was used to assess the significance of the differences between various groups of data. The differences were found to be significant: *—compared to control values, p < 0.01; x and xx—compared to the effect of H2O2 alone; x p < 0.01, xx p < 0.05, #—compared to the effect of α-T higher concentrations, p < 0.01.
Figure 1.
Shows that preincubation of immature brain cortical neurons with α-tocopherol (α-T) at nanomolar concentrations for 0.5 h prior to exposure of the cells to 0.2 mM H2O2 for 24 h did not increase the viability of brain cortical neurons (A); while preincubation with 10 and 100 nM α-T for 18 h prior to exposure of the neurons to 0.2 mM H2O2 for 24 h caused a pronounced increase in cortical neuron viability (B). H2O2 is designated as HP in the figure. Lactate dehydrogenase (LDH) method was used to determine neuron viability. The data bars (from the left to the right) in (A,B) show: (1) control values of LDH release from neurons; (2) control values of LDH release after incubation of the neurons with 100 µM α-T for 30 min (A) or for 18 h (B); (3) LDH release after neuron exposure to H2O2; (4) LDH release after preincubation of the neurons with 100 µM α-T prior to the cell exposure to H2O2; (5) LDH release after preincubation of the neurons with 100 nM α-T prior to the cell exposure to H2O2; (6) LDH release after preincubation of the neurons with 10 nM α-T prior to the cell exposure to H2O2. The results of one typical experiment from 9 experiments performed are shown as means ± SEM from 2–3 determinations in parallel samples. One way ANOVA followed by Tukey’s multiple comparison test was used to assess the significance of the differences between various groups of data. The differences were found to be significant: *—compared to control values, p < 0.01; x and xx—compared to the effect of H2O2 alone; x p < 0.01, xx p < 0.05, #—compared to the effect of α-T higher concentrations, p < 0.01.

Figure 2.
Shows that long preincubation with α-tocopherol (α-T) at nanomolar and micromolar concentrations diminished ROS accumulation in brain cortical neuron elicited by H2O2 to a great extent. The neurons were preincubated with α-T for 18 h. Then the fluorescent dye dichlorodihydrofluorescein diacetate was added to the incubation medium to a final concentration of 25 µM (see “Materials and Methods”). After 40 min incubation, the cells were exposed to 0.2 mM H2O2 for 4 h. H2O2 in the Figure is designated as HP. The data bars (from the left to the right) show: (1) control values; (2) control values after incubation of the neurons with 100 nM α-T; (3) control values after incubation of the neurons with 100 µM α-T; (4) ROS accumulation (arbitrary units) after neuron exposure to H2O2; (5) ROS accumulation after preincubation of the neurons with 100 µM α-T prior to the cell exposure to H2O2; (6) ROS accumulation after preincubation of the neurons with 100 nM α-T prior to the cell exposure to H2O2. The results of one typical experiment are shown as means ± SEM from 4–6 determinations in parallel samples. One-way ANOVA followed by Tukey’s test for multiple comparison was used to assess significance of the differences between various groups of data. The differences were found to be significant: *—compared to control values; x—compared to the effect of H2O2 alone; #—compared to the effect of 100 µM α-T (p < 0.01 in all cases).
Figure 2.
Shows that long preincubation with α-tocopherol (α-T) at nanomolar and micromolar concentrations diminished ROS accumulation in brain cortical neuron elicited by H2O2 to a great extent. The neurons were preincubated with α-T for 18 h. Then the fluorescent dye dichlorodihydrofluorescein diacetate was added to the incubation medium to a final concentration of 25 µM (see “Materials and Methods”). After 40 min incubation, the cells were exposed to 0.2 mM H2O2 for 4 h. H2O2 in the Figure is designated as HP. The data bars (from the left to the right) show: (1) control values; (2) control values after incubation of the neurons with 100 nM α-T; (3) control values after incubation of the neurons with 100 µM α-T; (4) ROS accumulation (arbitrary units) after neuron exposure to H2O2; (5) ROS accumulation after preincubation of the neurons with 100 µM α-T prior to the cell exposure to H2O2; (6) ROS accumulation after preincubation of the neurons with 100 nM α-T prior to the cell exposure to H2O2. The results of one typical experiment are shown as means ± SEM from 4–6 determinations in parallel samples. One-way ANOVA followed by Tukey’s test for multiple comparison was used to assess significance of the differences between various groups of data. The differences were found to be significant: *—compared to control values; x—compared to the effect of H2O2 alone; #—compared to the effect of 100 µM α-T (p < 0.01 in all cases).

Figure 3.
Shows the effect of incubation with 100 nM and 100 µM α-tocopherol (α-T) for 0.5, 1, 3, 7 and 24 h on the level of pAkt and total Akt in brain cortical neurons. Immunoblots obtained in one typical experiment from 6–7 experiments made are presented in (A). The data of 6–7 experiments made are presented as means ± SEM in (B). Red lines with squares show the effect of 100 nM α-T, black lines with rhombs the effect of 100 µM α-T. In this figure: *—the difference from the control level of pAkt is significant by paired Student’s t test after incubation with 100 nM α-T, p < 0.05.
Figure 3.
Shows the effect of incubation with 100 nM and 100 µM α-tocopherol (α-T) for 0.5, 1, 3, 7 and 24 h on the level of pAkt and total Akt in brain cortical neurons. Immunoblots obtained in one typical experiment from 6–7 experiments made are presented in (A). The data of 6–7 experiments made are presented as means ± SEM in (B). Red lines with squares show the effect of 100 nM α-T, black lines with rhombs the effect of 100 µM α-T. In this figure: *—the difference from the control level of pAkt is significant by paired Student’s t test after incubation with 100 nM α-T, p < 0.05.
Figure 4.
Shows that incubation of brain cortical neurons with 100 nM and 100 µM α-tocopherol (α-T) increased the level of pERK1/2 in these neurons. Immunoblots obtained in one typical experiment are presented in (A). The data are means ± SEM from 5–6 experiments in (B). Red lines with squares show the effect of 100 nM α-T, black lines with rhombs - the effect of 100 µM α-T. In this figure the difference is significant by paired Student’s t-test: *—between pERK1/2 level after exposure to 100 µM α-T and control pERK1/2 level, p < 0.05, x—between pERK1/2 level after exposure to 100 nM α-T and control pERK1/2 level, p < 0.05. The level of pERK1/2 significantly increased in brain cortical neurons after their exposure to 100 µM α-T for 1, 3, 7 and 24 h and to 100 nM α-T for 3 and 7 h. However, neither 100 nM, nor 100 µM α-T changed the total ERK1/2 level in brain cortical neurons, so it had no influence on the expression of this protein kinase.
Figure 4.
Shows that incubation of brain cortical neurons with 100 nM and 100 µM α-tocopherol (α-T) increased the level of pERK1/2 in these neurons. Immunoblots obtained in one typical experiment are presented in (A). The data are means ± SEM from 5–6 experiments in (B). Red lines with squares show the effect of 100 nM α-T, black lines with rhombs - the effect of 100 µM α-T. In this figure the difference is significant by paired Student’s t-test: *—between pERK1/2 level after exposure to 100 µM α-T and control pERK1/2 level, p < 0.05, x—between pERK1/2 level after exposure to 100 nM α-T and control pERK1/2 level, p < 0.05. The level of pERK1/2 significantly increased in brain cortical neurons after their exposure to 100 µM α-T for 1, 3, 7 and 24 h and to 100 nM α-T for 3 and 7 h. However, neither 100 nM, nor 100 µM α-T changed the total ERK1/2 level in brain cortical neurons, so it had no influence on the expression of this protein kinase.
Figure 5.
Shows the effect of preincubation of brain cortical neurons with 100 nM and 100 μM α-tocopherol (α-T) for 18 h prior to their exposure to 0.2 mM H2O2 for 24 h on pAkt and total Akt levels. Immunoblots obtained in one typical experiment are presented in (A). The results of 5–6 experiments are presented in (B) as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. In this figure: * and **—the differences are significant according to Student’s paired t test as compared to the level of pERK1/2 in brain cortical neurons exposed to H2O2 alone; *—the effect of preincubation with 100 nM α-T is significant, p < 0.05; **—the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.05.
Figure 5.
Shows the effect of preincubation of brain cortical neurons with 100 nM and 100 μM α-tocopherol (α-T) for 18 h prior to their exposure to 0.2 mM H2O2 for 24 h on pAkt and total Akt levels. Immunoblots obtained in one typical experiment are presented in (A). The results of 5–6 experiments are presented in (B) as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. In this figure: * and **—the differences are significant according to Student’s paired t test as compared to the level of pERK1/2 in brain cortical neurons exposed to H2O2 alone; *—the effect of preincubation with 100 nM α-T is significant, p < 0.05; **—the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.05.
Figure 6.
Shows the effect of preincubation of brain cortical neurons with 100 nM and 100 μM α-tocopherol (α-T) for 18 h prior to the cell exposure to 0.2 mM H2O2 for 24 h on pERK1/2 and total ERK1/2 levels. The immunoblots obtained in one typical experiment are presented in (A). The results of 5–6 experiments are presented in (B) as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. It is shown that 0.2 mM H2O2 activated ERK1/2 in brain cortical neurons (increased pERK1/2 level) 5 min after its application, then ERK1/2 remained at the same high level during 24 h of prooxidant action. However, if these neurons were preincubated with 100 nM and 100 μM α-T for 18 h and then exposed to 0.2 mM H2O2 for 24 h, the activity of ERK1/2 was not high, it was close to control values 12 and 24 h after exposure of the neurons to this prooxidant, the effect of preincubation with 100 nM and 100 µM α-T was significant. Preincubation with α-T (100 µM) caused a significant increase of the pERK1/2 level as compared to the effect of H2O2 alone at early stages of its action—5 min after cell exposure to H2O2, but 100 nM α-T did not exert such an effect. No change in the total ERK1/2 level was revealed as a result of the exposure of neurons to H2O2 alone or to H2O2 after preincubation with α-T, which means that the expression of this protein kinase was not changed. In this figure: * and **—the differences are significant according to Student’s paired t test as compared to the level of pERK1/2 in brain cortical neurons exposed to H2O2 alone; *—the effect of preincubation with 100 µM α-T is significant; p < 0.05, **—the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.05.
Figure 6.
Shows the effect of preincubation of brain cortical neurons with 100 nM and 100 μM α-tocopherol (α-T) for 18 h prior to the cell exposure to 0.2 mM H2O2 for 24 h on pERK1/2 and total ERK1/2 levels. The immunoblots obtained in one typical experiment are presented in (A). The results of 5–6 experiments are presented in (B) as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. It is shown that 0.2 mM H2O2 activated ERK1/2 in brain cortical neurons (increased pERK1/2 level) 5 min after its application, then ERK1/2 remained at the same high level during 24 h of prooxidant action. However, if these neurons were preincubated with 100 nM and 100 μM α-T for 18 h and then exposed to 0.2 mM H2O2 for 24 h, the activity of ERK1/2 was not high, it was close to control values 12 and 24 h after exposure of the neurons to this prooxidant, the effect of preincubation with 100 nM and 100 µM α-T was significant. Preincubation with α-T (100 µM) caused a significant increase of the pERK1/2 level as compared to the effect of H2O2 alone at early stages of its action—5 min after cell exposure to H2O2, but 100 nM α-T did not exert such an effect. No change in the total ERK1/2 level was revealed as a result of the exposure of neurons to H2O2 alone or to H2O2 after preincubation with α-T, which means that the expression of this protein kinase was not changed. In this figure: * and **—the differences are significant according to Student’s paired t test as compared to the level of pERK1/2 in brain cortical neurons exposed to H2O2 alone; *—the effect of preincubation with 100 µM α-T is significant; p < 0.05, **—the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.05.

Figure 7.
Shows the effect of H
2O
2 and preincubation with α-tocopherol (α-T) on the level of the active fragment of PKCδ with molecular mass 40 kDa and the level of total PKCδ in brain cortical neurons. These neurons were preincubated with 100 nM and 100 μM α-T (or without it) for 18 h and then exposed to 0.2 mM H
2O
2 for 24 h. Immunoblots obtained in one typical experiment (from 5 experiments made) show that H
2O
2 increased the level of catalytically active 40 kDa fragment of PKCδ in neurons up to 12 h after the beginning of their exposure to this prooxidant. It means that H
2O
2 activated PKCδ in brain cortical neurons. However, preincubation with 100 nM and 100 μM α-T diminished the increase of the level of 40 kDa fragment of PKCδ induced by H
2O
2. H
2O
2 and α-T had no effect on total PKCδ level (
Figure 7), which means that they did not change the expression of this enzyme.
Figure 7.
Shows the effect of H
2O
2 and preincubation with α-tocopherol (α-T) on the level of the active fragment of PKCδ with molecular mass 40 kDa and the level of total PKCδ in brain cortical neurons. These neurons were preincubated with 100 nM and 100 μM α-T (or without it) for 18 h and then exposed to 0.2 mM H
2O
2 for 24 h. Immunoblots obtained in one typical experiment (from 5 experiments made) show that H
2O
2 increased the level of catalytically active 40 kDa fragment of PKCδ in neurons up to 12 h after the beginning of their exposure to this prooxidant. It means that H
2O
2 activated PKCδ in brain cortical neurons. However, preincubation with 100 nM and 100 μM α-T diminished the increase of the level of 40 kDa fragment of PKCδ induced by H
2O
2. H
2O
2 and α-T had no effect on total PKCδ level (
Figure 7), which means that they did not change the expression of this enzyme.
Figure 8.
Shows the effect of incubation of brain cortical neurons with α-tocopherol (α-T) for 18 h on the basal Bax/Bcl-2 ratio. This ratio was taken as 100% in control brain cortical neurons. The data of five experiments made are presented as means ± SEM. α-T (100 nM and 100 µM) was shown to decrease the basal Bax/Bcl-2 ratio in brain cortical neurons. The diminution of this ratio was not pronounced, but it was significant. *—the difference is significant according to Student’s paired t test as compared to the Bax/Bcl-2 ratio in control cortical neurons, p < 0.02.
Figure 8.
Shows the effect of incubation of brain cortical neurons with α-tocopherol (α-T) for 18 h on the basal Bax/Bcl-2 ratio. This ratio was taken as 100% in control brain cortical neurons. The data of five experiments made are presented as means ± SEM. α-T (100 nM and 100 µM) was shown to decrease the basal Bax/Bcl-2 ratio in brain cortical neurons. The diminution of this ratio was not pronounced, but it was significant. *—the difference is significant according to Student’s paired t test as compared to the Bax/Bcl-2 ratio in control cortical neurons, p < 0.02.
Figure 9.
Shows the effect of exposure of cortical neurons to 0.2 mM H2O2 for 24 h and of preincubation with 100 nM and 100 μM α-tocopherol (α-T) for 18 h on the level of Bcl-2 and Bax in brain cortical neurons. The results of immunoblotting obtained in one typical experiment are shown in (A). The results of 6–7 experiments are shown in (B,C), respectively, as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. * and **—the differences are significant according to Student’s paired t test as compared to the level of Bcl-2 in brain cortical neurons exposed to H2O2 alone, * the effect of preincubation with 100 nM α-T is significant, * p < 0.05, ** the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.02.
Figure 9.
Shows the effect of exposure of cortical neurons to 0.2 mM H2O2 for 24 h and of preincubation with 100 nM and 100 μM α-tocopherol (α-T) for 18 h on the level of Bcl-2 and Bax in brain cortical neurons. The results of immunoblotting obtained in one typical experiment are shown in (A). The results of 6–7 experiments are shown in (B,C), respectively, as means ± SEM. Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. * and **—the differences are significant according to Student’s paired t test as compared to the level of Bcl-2 in brain cortical neurons exposed to H2O2 alone, * the effect of preincubation with 100 nM α-T is significant, * p < 0.05, ** the effect of preincubation with both 100 nM and 100 µM α-T is significant, p < 0.02.
Figure 10.
Shows the effect of H2O2 and preincubation with α-tocopherol (α-T) on the Bax/Bcl-2 ratio in brain cortical neurons. The levels of Bcl-2 and Bax 3, 5, 12 and 24 h after brain cortical neuron exposure to 0.2 mM H2O2 after preincubation for 18 h with 100 nM α-T and 100 μM α-T (or without it) are shown in (A). The results of five experiments on the Bax/Bcl-2 ratio in brain cortical neurons are presented as means ± SEM in (B). Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. * and **—the differences are significant according to Student’s paired t test as compared to the initial level of Bax/Bcl-2 ratio (0 point) and to the level of this ratio in brain cortical neurons exposed to H2O2 after preincubation with 100 nM and 100 μM α-T, * p < 0.05, ** p < 0.02. It means that the effect of preincubation with both 100 nM and 100 µM α-T is significant.
Figure 10.
Shows the effect of H2O2 and preincubation with α-tocopherol (α-T) on the Bax/Bcl-2 ratio in brain cortical neurons. The levels of Bcl-2 and Bax 3, 5, 12 and 24 h after brain cortical neuron exposure to 0.2 mM H2O2 after preincubation for 18 h with 100 nM α-T and 100 μM α-T (or without it) are shown in (A). The results of five experiments on the Bax/Bcl-2 ratio in brain cortical neurons are presented as means ± SEM in (B). Red lines with squares show the effect of H2O2 alone, black lines with rhombs—effect of H2O2 after preincubation with 100 nM α-T, green lines with triangles—effect of H2O2 after preincubation with 100 μM α-T. * and **—the differences are significant according to Student’s paired t test as compared to the initial level of Bax/Bcl-2 ratio (0 point) and to the level of this ratio in brain cortical neurons exposed to H2O2 after preincubation with 100 nM and 100 μM α-T, * p < 0.05, ** p < 0.02. It means that the effect of preincubation with both 100 nM and 100 µM α-T is significant.

Table 1.
Shows the protective effect of preincubation with α-T for 18 h prior to brain cortical neuron exposure to 0.2 mM H2O2 for 24 h expressed as rescue rates of α-T. Cell viability was assessed by the LDH method. The data are means ± SEM from 7–9 experiments. The difference in the LDH activity released from cortical neurons exposed to H2O2 in the absence and presence of α-T was determined. The ratio of this difference to the increase of LDH activity released from neurons to the medium in the presence of H2O2 alone (taken as 100%) corresponded to the rescue rates of α-T against H2O2-induced cell death. The formula is ([LDH release in H2O2 − LDH release in H2O2 and α-T]/[LDH release in H2O2 − LDH release in control]) x 100. In this table: *—the protective effect of α-T is significant, p < 0.01; x and #—the differences are significant according to Student’s t test as compared to the effect of α-T at lower concentrations, x p < 0.02, # p < 0.01.
Table 1.
Shows the protective effect of preincubation with α-T for 18 h prior to brain cortical neuron exposure to 0.2 mM H2O2 for 24 h expressed as rescue rates of α-T. Cell viability was assessed by the LDH method. The data are means ± SEM from 7–9 experiments. The difference in the LDH activity released from cortical neurons exposed to H2O2 in the absence and presence of α-T was determined. The ratio of this difference to the increase of LDH activity released from neurons to the medium in the presence of H2O2 alone (taken as 100%) corresponded to the rescue rates of α-T against H2O2-induced cell death. The formula is ([LDH release in H2O2 − LDH release in H2O2 and α-T]/[LDH release in H2O2 − LDH release in control]) x 100. In this table: *—the protective effect of α-T is significant, p < 0.01; x and #—the differences are significant according to Student’s t test as compared to the effect of α-T at lower concentrations, x p < 0.02, # p < 0.01.
α-T Concentration | 100 μM α-T | 10 μM α-T | 1 μM α-T | 100 nM α-T | 10 nM α-T | 1 nM α-T |
---|
Rescue rates (%) | 64.3 ± 7.2 * | 67.4 ± 11.9 * | 60.0 ± 11.4 * | 52.5 ± 7.4 *x | 27.3 ± 5.1 *# | 5.1 ± 2.9 |
Table 2.
Shows that the rescue rates of α-T against H2O2-induced brain cortical neuron death were significantly lower in the presence of an inhibitor of PI 3-kinase (LY294002), an inhibitor of MEK1/2 (SL327) and an inhibitor of PKCδ (rottlerin) than in their absence in the incubation medium. Cell viability was assessed by the LDH method. The data are means ± SEM from 5–6 experiments. Preincubation with protein kinase inhibitors was performed for 0.5 h, then α-T was added for 18 h before brain cortical neuron exposure to 0.2 mM H2O2 for 24 h. In this table: ** and *—the protective effect of α-T is significant, ** p < 0.02, * p < 0.05; x and #—the differences are significant as compared to the effect of α-T in the absence of inhibitors by paired Student’s t-test, x p < 0.02, # p < 0.05.
Table 2.
Shows that the rescue rates of α-T against H2O2-induced brain cortical neuron death were significantly lower in the presence of an inhibitor of PI 3-kinase (LY294002), an inhibitor of MEK1/2 (SL327) and an inhibitor of PKCδ (rottlerin) than in their absence in the incubation medium. Cell viability was assessed by the LDH method. The data are means ± SEM from 5–6 experiments. Preincubation with protein kinase inhibitors was performed for 0.5 h, then α-T was added for 18 h before brain cortical neuron exposure to 0.2 mM H2O2 for 24 h. In this table: ** and *—the protective effect of α-T is significant, ** p < 0.02, * p < 0.05; x and #—the differences are significant as compared to the effect of α-T in the absence of inhibitors by paired Student’s t-test, x p < 0.02, # p < 0.05.
Sample | Rescue Rates of α-T, % | Sample | Rescue Rates of α-T, % |
---|
100 nM α-T | 52.4 ± 13.1 ** | 100 μM α-T | 63.35 ± 12.6 ** |
100 nM α-T + 10 µM SL327 | 31.35 ± 11.1 *,# | 100 μM α-T + 10 μM SL327 | 36.9 ± 12.3 * |
100 nM α-T + 50 µM LY294002 | 10.1 ± 4.6 x | 100 μM α-T + 50 μM LY294002 | 27.6 ± 14.7 # |
100 nM α-T | 50.0 ± 7.5 ** | 100 μM α-T | 52.2 ± 6.4 ** |
100 nM α-T + 5 µM rottlerin | 20.9 ± 4.6 **,x | 100 μM α-T + 5 μM rottlerin | 33.6 ± 4.8 **,x |