Suppression of Lipid Accumulation in the Differentiation of 3T3-L1 Preadipocytes and Human Adipose Stem Cells into Adipocytes by TAK-715, a Specific Inhibitor of p38 MAPK
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Materials
2.2. 3T3-L1 Cells or hASCs Culture and Differentiation
2.3. Oil Red O Staining
2.4. Cell Count Analysis
2.5. Measurement of Intracellular TG Content
2.6. Preparation of Whole-Cell Lysates
2.7. Westerm Blotting Analysis
2.8. RT-qPCR
2.9. Statistical Analysis
3. Results
3.1. TAK-715 at 10 μM Markedly Suppresses Intracelullar Lipid Accumulation and TG Content during the Adipogenesis of 3T3-L1 Preadipocyte
3.2. TAK-715 Significantly Reduces the Expression and Phosphorylation Levels of C/EBP-α, PPAR-γ, and STAT-3 in 3T3-L1 Cells Following Differentiation
3.3. TAK-715 Significantly Lower the Expressions of FAS and Perilipin A during Preadipocyte Differentiation in 3T3-L1 Cells
3.4. TAK-715 Significantly Reduces the Phosphorylation Levels of ATF-2, a p38 MAPK Downstream Effector, in Differentiating 3T3-L1 Preadipocyte
3.5. TAK-715 Markedly Reduces Lipid Formation and p38 MAPK Phosphorylation in hASCs Adipocyte Differentiation
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibodies | Dilution Used | Source | Catalog No. |
---|---|---|---|
Primary Antibodies | |||
C/EBP-α | 1:2000 | Santa Cruz Biotechnology | sc-61 |
PPAR-γ | 1:2000 | Santa Cruz Biotechnology | sc-7272 |
p-STAT-3 | 1:2000 | Santa Cruz Biotechnology | sc-8059 |
T-STAT-3 | 1:2000 | Santa Cruz Biotechnology | sc-8019 |
p-STAT-5 | 1:2000 | Santa Cruz Biotechnology | sc-10180b |
T-STAT-5 | 1:2000 | Santa Cruz Biotechnology | sc-835 |
FAS | 1:2000 | BD Bioscience | #9452 |
Perilipin A | 1:2000 | BioVision | #3948-200 |
p-ATF-2 | 1:2000 | Cell Signaling | #9225 |
T-ATF-2 | 1:2000 | Cell Signaling | #9226 |
p-p38 MAPK | 1:2000 | Cell Signaling | #9211 |
T-p38 MAPK | 1:2000 | Cell Signaling | #9212 |
β-actin | 1:10,000 | Sigma | A5441 |
Gene | Forward | Reverse |
---|---|---|
FAS | TTGCTGGCACTACAGAATGC | AACAGCCTCAGAGCGACAAT |
Perilipin A | CTTTCTCGACACACCATGGAAAC | CCACGTTATCCGTAACACCCTTCA |
18S rRNA | GGTGAAGGTCGGTGTGAACG | GGTAGGAACACGGAAGGCCA |
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Perumal, N.L.; Mufida, A.; Yadav, A.K.; Son, D.-G.; Ryoo, Y.-W.; Kim, S.-A.; Jang, B.-C. Suppression of Lipid Accumulation in the Differentiation of 3T3-L1 Preadipocytes and Human Adipose Stem Cells into Adipocytes by TAK-715, a Specific Inhibitor of p38 MAPK. Life 2023, 13, 412. https://doi.org/10.3390/life13020412
Perumal NL, Mufida A, Yadav AK, Son D-G, Ryoo Y-W, Kim S-A, Jang B-C. Suppression of Lipid Accumulation in the Differentiation of 3T3-L1 Preadipocytes and Human Adipose Stem Cells into Adipocytes by TAK-715, a Specific Inhibitor of p38 MAPK. Life. 2023; 13(2):412. https://doi.org/10.3390/life13020412
Chicago/Turabian StylePerumal, Nivethasri Lakshmana, Amila Mufida, Anil Kumar Yadav, Dae-Gu Son, Young-Wook Ryoo, Sung-Ae Kim, and Byeong-Churl Jang. 2023. "Suppression of Lipid Accumulation in the Differentiation of 3T3-L1 Preadipocytes and Human Adipose Stem Cells into Adipocytes by TAK-715, a Specific Inhibitor of p38 MAPK" Life 13, no. 2: 412. https://doi.org/10.3390/life13020412