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. 2021 Oct 6;10(20):e022568. doi: 10.1161/JAHA.121.022568

Figure 5. Regulation of transforming growth factor‐β1 and collagen I expression by Rho/Rho‐associated coiled‐coil containing kinase (ROCK) isoforms.

Figure 5

A, Protein and (B) mRNA levels of transforming growth factor‐β1 and collagen I in wild‐type primary aortic smooth muscle cells (SMC) treated with Ang II (10 μmol/L) or Y‐27632 (20 μmol/L) for 24 hours. n=4 cell isolates in each group. C, Representative immunoblots and densitometric quantification of transforming growth factor‐β1 and collagen I in wild‐type SMC treated with control (scrambled), Rock1 or Rock2 small interfering RNA. After transfection of small interfering RNA for 48 hours, SMC were stimulated with or without Ang II (10 μmol/L) for 24 hours, n=4 cell isolates in each group. D, Representative immunoblots and densitometric quantification of connective tissue growth factor and collagen I in wild‐type, Rock1 −/− or Rock2 −/− mouse embryonic fibroblasts treated with transforming growth factor‐β1 (10 ng/mL) for 24 hours; n=4 cell isolates in each group. Results are expressed as mean±SEM. P values were calculated using 1‐way ANOVA with Tukey Honest Significant Difference test. Ang II indicates angiotensin II; CTGF, connective tissue growth factor; p‐MBS indicates phosphorylated myosin‐binding subunit; ROCK/Rock, Rho/Rho‐associated coiled‐coil containing kinase; siRNA, small interfering RNA; TGF, transforming growth factor; t‐MBS, total myosin‐binding subunit; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; Hprt, hypoxanthine phosphoribosyltransferase; Col1a1, collagen type I alpha 1 chain; and WT, wild‐type.