FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Human ESC Culture and Differentiation
2.2. Gene and Expression Analysis
2.3. Immunofluorescence
2.4. SHH-ELISA
2.5. SHH Activity Assay
2.6. Transcriptomics
2.7. Proteomics
2.8. Western Blot
2.9. Statistics
3. Results
3.1. Effect of Growth Factors on Early Pancreatic Development
3.2. Small Molecule Inhibition of FGF-Signaling
3.3. Secretome Analysis
3.4. FGF-Signaling through FGFR1c/3c
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene Names | Protein Names | Mean LFQ (d0) | Mean LFQ (d4) | Fold Change LFQ (d4/d0) | Fold Change Transcription (d4/d0) |
---|---|---|---|---|---|
FGF17 | Fibroblast growth factor 17 | 2.3 × 106 | 2.2 × 109 | 918.56 | 418.27 |
SFRP1 | Secreted frizzled-related protein 1 | 4.0 × 106 | 1.6 × 109 | 392.12 | 2.43 |
FGF8 | Fibroblast growth factor 8 | 1.8 × 106 | 6.2 × 108 | 338.51 | 64.36 |
CER1 | Cerberus | 9.6 × 107 | 2.1 × 1010 | 223.01 | 816.30 |
SFRP3 (FRZB) | Secreted frizzled-related protein 3 | 1.6 × 106 | 2.9 × 108 | 185.39 | 46.72 |
DKK1 | Dickkopf-related protein 1 | 1.3 × 106 | 1.7 × 108 | 129.79 | 27.65 |
FLRT3 | Leucine-rich repeat transmembrane protein FLRT3 | 2.6 × 106 | 3.0 × 108 | 117.66 | 38.37 |
STC1 | Stanniocalcin-1 | 2.9 ×107 | 2.5 × 109 | 85.60 | 18.76 |
NPPB | Natriuretic peptides | 2.5 × 106 | 1.5 × 108 | 60.52 | 890.54 |
MDK | Midkine | 1.7 × 107 | 7.6 × 108 | 43.79 | 2.77 |
INHBA | Inhibin beta A chain | 6.0 × 106 | 2.6 × 108 | 43.67 | 0.02 |
BMP2 | Bone morphogenetic protein 2 | 1.8 × 106 | 6.9 × 107 | 38.04 | 116.29 |
CPE | Carboxypeptidase E | 9.5 × 107 | 2.8 × 109 | 29.74 | 15.76 |
LEFTY2 | Left-right determination factor 2 | 8.9 × 108 | 2.0× 1010 | 23.03 | 1757.26 |
LEFTY1 | Left-right determination factor 1 | 9.2 × 108 | 2.0 × 1010 | 21.16 | 679.87 |
GRN | Granulins | 1.0 × 107 | 1.7 × 108 | 17.12 | 1.54 |
BMP7 | Bone morphogenetic protein 7 | 3.0 × 106 | 4.4 × 107 | 14.70 | 3.16 |
NRP2 | Neuropilin-2 | 1.1 × 107 | 1.1 × 108 | 9.73 | 0.40 |
ROR2 | Tyrosine-protein kinase transmembrane receptor ROR2 | 1.3 ×107 | 9.8 × 107 | 7.31 | 19.48 |
ITGB5 | Integrin beta-5 | 1.1 × 107 | 7.4 ×107 | 6.71 | 2.81 |
HSPB1 | Heat shock protein beta-1 | 5.9 × 107 | 2.7 × 108 | 4.54 | 2.00 |
PTK7 | Inactive tyrosine-protein kinase 7 | 2.9 × 108 | 1.3 × 109 | 4.54 | 1.10 |
NXN | Nucleoredoxin | 5.7 ×107 | 2.1 × 108 | 3.64 | 1.70 |
GPC4 | Glypican-4 | 6.7 × 108 | 2.4 × 109 | 3.63 | 1.61 |
NCBP1 | Nuclear cap-binding protein subunit 1 | 2.5 × 107 | 9.1× 107 | 3.59 | 0.81 |
SNW1 | SNW domain-containing protein 1 | 1.5 × 106 | 5.0 × 106 | 3.25 | 0.83 |
FKBP8 | Peptidyl-prolyl cis-trans isomerase FKBP8 | 4.9 × 106 | 1.4 × 107 | 2.93 | 0.76 |
GTF2F2 | General transcription factor IIF subunit 2 | 8.6 × 106 | 1.9 × 107 | 2.22 | 0.87 |
ADAM10 | Disintegrin and metalloproteinase domain-containing protein 10 | 7.2 × 107 | 1.5 × 108 | 2.11 | 1.36 |
TIAL1 | Nucleolysin TIAR | 1.9 × 108 | 1.1 × 108 | 0.57 | 0.82 |
DDB1 | DNA damage-binding protein 1 | 1.6 × 109 | 7.4 × 108 | 0.45 | 0.95 |
CTGF | Connective tissue growth factor | 1.2 × 109 | 4.7 × 108 | 0.392 | 0.39 |
WDR12 | Ribosome biogenesis protein WDR12 | 1.5 × 108 | 4.3 × 107 | 0.290 | 0.53 |
PARP1 | Poly [ADP-ribose] polymerase 1 | 3.0 × 109 | 5.8 × 108 | 0.191 | 0.51 |
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Dettmer, R.; Cirksena, K.; Münchhoff, J.; Kresse, J.; Diekmann, U.; Niwolik, I.; Buettner, F.F.R.; Naujok, O. FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells. Cells 2020, 9, 1927. https://doi.org/10.3390/cells9091927
Dettmer R, Cirksena K, Münchhoff J, Kresse J, Diekmann U, Niwolik I, Buettner FFR, Naujok O. FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells. Cells. 2020; 9(9):1927. https://doi.org/10.3390/cells9091927
Chicago/Turabian StyleDettmer, Rabea, Karsten Cirksena, Julia Münchhoff, Jasmin Kresse, Ulf Diekmann, Isabell Niwolik, Falk F. R. Buettner, and Ortwin Naujok. 2020. "FGF2 Inhibits Early Pancreatic Lineage Specification during Differentiation of Human Embryonic Stem Cells" Cells 9, no. 9: 1927. https://doi.org/10.3390/cells9091927