Figure 4. A shows the induction of BRM mRNA following the gene-specific shRNA-mediated knockdown of HDAC3, HDAC9, GATA3 or MEF2D in G401 and KD cell lines, which resulted in a greater than > 5-fold induction for each gene in either cell line.
B shows the G401 and KD cell lines harboring either scrambled shRNA (scr-shRNA) or anti-BRM shRNA; these cell lines were then subjected to gene-specific shRNA-mediated knockdown of HDAC3, HDAC9, MEF2D or GATA3. The knockdown of HDAC3, HDAC9, MEF2D or GATA3 displayed a statistically significant degree of growth inhibition in the cell lines harboring the scrambled shRNA (65-80%) in comparison to the daughter cell lines harboring the antiBRM shRNA (15-30%; p<0.05). C illustrates the level of HDAC9 mRNA in 11 Rhabdoid cell lines. Four previously characterized non-Rhabdoid cell lines, H441, H460 (BRM-positive:low HDAC9), and SW13 and C33A (BRM-negative: high HDAC9) were used as negative and positive controls, respectively, for HDAC9. The level of HDAC9 mRNA is approximately the same as in the BRM-negative non-Rhabdoid cell lines, SW13 and C33A, and on average is ~47±13 fold higher than the HDAC9 mRNA level observed in the BRM-positive Rhabdoid cell line, TTC642. D demonstrates the change in HDAC9 mRNA level following the gene-specific shRNA-mediated knockdown of either GATA3 or MEF2D. MEF2D knockdowns caused 15- and 16-fold down-regulation of HDAC9 in G401 and KD cells, respectively, compared to the cells harboring the scr-shRNA. GATA3 knockdowns resulted in a 75- and 256-fold down-regulation of HDAC9 in G401 and KD cells, respectively.