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. 2015 Oct 8;8(1):73–87. doi: 10.1093/jmcb/mjv059

Figure 4.

Figure 4

Characterization of murine Smad1/5-null aMABs and eMABs. (A) IF analysis for P-SMAD1/5/8 (red) and GFP (green) in Smad1/5-null and WT aMABs. (B) WB analysis (left panel) and quantification (right panel) of P-SMAD1/5/8 and SMAD8 levels normalized by GAPDH in Smad1/5-null aMABs. (C) RT-qPCR analysis of Smad8 mRNA levels in Smad1/5-null aMABs. Data are expressed as fold induction compared with WT aMABs and normalized to steady-state mRNA levels of Gapdh, Hprt, and Tbp housekeeping genes. (D) IF analysis for P-SMAD1/5/8 (red) and GFP (green) in Smad1/5-null and WT eMABs. (E) WB analysis (left panel) and quantification (right panel) of P-SMAD1/5 and SMAD8 levels normalized by GAPDH in Smad1/5-null eMABs. (F and G) IF analysis for MyHC (red) and GFP (green) in Smad1/5-null aMABs (F) and eMABs (G) co-cultured with C2C12 (1:1) and subjected to myogenic differentiation for 5 days. (H) Fusion index of chimeric myotubes generated from Smad1/5-null aMABs (F) or eMABs (G) in co-culture experiments is reported as percentage with respect to WT MABs. *P < 0.05, ***P < 0.001 vs. WT. Scale bar, 500 μm.