Figure 3. Pask is required for myogenic conversion of C3H10T1/2 cells by MyoD.
(A) Schematic depiction of the mechanism by which MyoD-induces the myogenic conversion of adipogenic C3H10T1/2 cells. (B) qRT-PCR analysis of the indicated mRNAs in C3H10T1/2 cells expressing MyoD in the presence of DMSO or 25 µM BioE-1197 during differentiation. n = 3, Error bars ± S.D ***p<0.0005, p<0.005. (C) MyoD-expressing C3H10T1/2 cells were allowed to differentiate in the presence of DMSO or 25 µM BioE-1197 and processed for immunofluorescence microscopy using anti-MyoG antibody. Scale bar = 20 µm. (D) Quantification of MyoD+ and MyoG+ cells in the presence of DMSO or BioE-1197 from (C). n = 3 independent experiments each with 100 cells counted. Error bars ± S.D. **p<0.005. (E) Fusion index on Day 1 and 2 of differentiation for C3H10T1/2 cells expressing MyoD in the presence of DMSO or 25 µM BioE-1197. Inset shows representative MHC staining on Day 2 of differentiation. Scale bar = 20 µm. (F) qRT-PCR analysis of endogenous mouse Mylpf and Myog and rat Myog upon GFP or rat MyoG expression in MyoD-expressing C3H10T1/2 cells treated with either DMSO or 25 µM BioE-1197. n = 3, Error bars ± S.D. ***p<0.0005, **p<0.005.
DOI: http://dx.doi.org/10.7554/eLife.17985.013
Figure 3—figure supplement 1. Pask is required for myogenic conversion of mesenchymal stem cells by MyoD.
