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. 2015 Jul 1;16(8):1037–1050. doi: 10.15252/embr.201540159

Figure 1.

Figure 1

Brm and Brg1 show specific profiles of expression and activities during skeletal muscle differentiation

  1. Time course of protein expression during terminal differentiation of C2C12 myoblasts representative of three independent experiments. Myoblasts were cultured in growth medium (GM) until they reached confluence, and then shifted to differentiate in differentiation medium (DM) for 48 h. Cellular extracts were analyzed by Western blot with antibodies against BRG1, Brm, myosin heavy chain (MyHC), and cyclin D1. Cdk4 probing was used to check for equal loading of the samples.
  2. Immunofluorescence analysis of Brm and Brg1 expression in C2C12 cells cultured in GM or DM conditions. Scale bar, 50 μm.
  3. Efficiency of BRM and BRG1 knockdown at 48 h post-transfection performed in C2C12 cells using siRNAs (control interference is a scrambled sequence and referred as siScr) was monitored by qRT–PCR. Data are presented as average ± SEM (n > 3).
  4. Immunofluorescence for Brm or Brg1 performed in proliferating myoblasts upon siRNA against Brg1 (siBrg1), Brm (siBrm) or scrambled (siScr) to check for efficient depletion of the proteins. Scale bar, 50 μm.
  5. Brightfield images and MyHC staining were performed at various time points of differentiation in C2C12 cells in which siRNAs were delivered in GM as depicted in the scheme above. Scale bar, 50 μm.
  6. Quantification of fusion index of three independent experiments calculated as percentage of nuclei within MyHC-expressing myotubes. Data are presented as average ± SEM (n > 3). *P < 0.05; ***P < 0.001 (unpaired Student’s t-test).