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. 2021 Oct 26;13(21):5373. doi: 10.3390/cancers13215373

Figure 2.

Figure 2

Transcriptional regulation of the newly identified DLC1-dependent genes. (A) mRNA expression of TSPAN5, VCAN and CDH2 in HepG2 cells transiently transfected with ctrl siRNA (sictrl), DLC1 siRNA (siDLC1) and a combination of DLC1 siRNA (siDLC1) and MRTF-A/-B siRNA (siMRTF-A/-B) or a combination of DLC1 siRNA (siDLC1) and FLNA siRNA (siFLNA) was assessed by qRT-PCR. The respective gene-specific primers were used and normalization to the 18 S rRNA was carried out. Values are means ± SD (n = 3); **, p < 0.01 and ***, p < 0.001. (B) Expression of MRTF-A, VCAN and TSPAN5 mRNA in HuH7 cells after RNAi-mediated MRTF-A/-B knockdown (siMRTF-A/-B), followed by ectopic expression of MRTF-A-N100 for 3 days, determined by qRT-PCR as described above. Values are means ± SD (n = 3); *, p < 0.05, **, p < 0.01 and ***, p < 0.001. (C) ChIP was performed in three independent chromatin preparations from FLNA-expressing A7 cells compared to FLNA-deficient M2 cells with MRTF-A and IgG antibodies for pulldown. The specific primers for VCAN, TSPAN5 and CDH2 promoters listed in Table S7 were used. Values are means ± SD (n = 3); *, p < 0.05 and **, p < 0.01.