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. 2020 Sep 4;8:839. doi: 10.3389/fcell.2020.00839

FIGURE 6.

FIGURE 6

Sp1-mediated phosphodiesterase 5 (PDE5) transcription is essential for myocardin-related transcription factor A (MRTF-A) to mediate Ang-II-induced prohypertrophic response in cardiomyocytes. (A) H9C2 cells were transfected with siRNA targeting MRTF-A or scrambled siRNA (SCR) in the presence or absence of ectopic Sp1 followed by treatment with Ang II (1 μM) for 24 h. Gene expression was examined by quantitative PCR (qPCR). (B) H9C2 cells were treated with Ang II (1 μM) and CCG-1423 (10 μM) in the presence or absence of ectopic Sp1 for 24 h. Gene expression was examined by qPCR. (C) H9C2 cells were transfected with siRNA targeting MRTF-A or scrambled siRNA (SCR) in the presence or absence of ectopic Sp1 followed by treatment with Ang II (1 μM) for 24 h. Immunofluorescence staining was performed with anti-α-actinin. Cross-sectional areas were quantified by Image Pro. (D) H9C2 cells were treated with Ang II (1 μM) and CCG-1423 (10 μM) in the presence or absence of ectopic Sp1 for 24 h. Immunofluorescence staining was performed with anti-α-actinin. (E) Primary neonatal rat ventricular myocytes were transfected with siRNA targeting MRTF-A or scrambled siRNA (SCR) followed by treatment with Ang II (1 μM) for 24 h. Gene expression was examined by qPCR. (F) Primary neonatal rat ventricular myocytes were treated with Ang II (1 μM) and CCG-1423 (10 μM) for 24 h. Gene expression was examined by qPCR. (G) A schematic model. Data represent averages of three independent experiments, and error bars represent SEM. *p < 0.05; **p < 0.01; ***p < 0.001. Scale bar: 25 μm.