Figure 4. Silencing JARID2 results in myogenic differentiation and TPA-induced differentiation can be reversed by increasing JARID2 levels in RMS cells.
(A) Phase contrast image of RH30 cells with and without JARID2 siRNA silencing 72 hours post transfection (scale bar, 100 microns). Arrows show elongated cells consistent with myogenic differentiation. (B) Myogenin (MYOG) and Myosin Light Chain (MYL1) expression in RH30 and RD by Western blotting 72hrs after transfection with siRNAa against JARID2. (C) JARID2 expression is reduced in TPA-induced differentiation of RMS cell lines. MYOG and JARID2 mRNA expression in RD cells at 72hrs post TPA addition relative to DMSO control. Dark bars represent expression of MYOG in TPA-induced and DMSO control samples. Light grey bars represent JARID2 expression in these samples. Error bars represent SD of 3 replicates. (D) JARID2 overexpression in the RD cell line reduced expression of MYL1 expression in TPA-induced cells relative to DMSO vector-only control (pCI-Neo). Error bars represent SD of 3 replicates. (E) Confirmation of rescue of myogenic marker expression with overexpression of JARID2 in the RD cell line by Western blot. JARID2 overexpression in the RD cell line reduces expression of MYOG and MYL1 in TPA-induced cells relative to DMSO vector-only control (pCI-Neo).