Figure 2.

Relative levels of MEF2Cγ- transcripts correlated with the abundance of RBM4. (A) Mock vector-transfected C3H10T1/2 cells were cultured in growth (GM) or differentiating medium (DM) for 7 days. Cells overexpressing FLAG-RBM4 or its truncated isoform, Iso2, were cultured in GM for 24 h. An RT-PCR analysis was performed as described in Fig. 1. Extracted total lysates from the same batch of cells were subjected to an immunoblotting assay by probing with the indicated antibodies. (B) C3H10T1/2 cells were transiently transfected with a mock vector, or a FLAG-RBM4-expressing vector, or a RBM4-targeting shRNA expressing vector and cultured in GM for 24 h. RT-PCR and immunoblotting assays were performed as described in panel A. Total RNA was extracted from cells overexpressing (C) wild-type or mutant FLAG-RBM4 proteins (D) or distinct FLAG-tagged RNA-binding proteins. The RT-PCR and immunoblotting assay were performed as described in the previous panel. The bar graph showed the relative ratio of MEF2Cγ- over total MEF2C transcripts using TotalLab Quant Software (*p < 0.05; **p < 0.01; ***p < 0.005).