Fig. 3

Rhein attenuated Ang II-induced CF activation in vitro. A The 24-h exposure of CFs to rhein (0, 5, 10, 25, 50, and 100 μM). Cellular viability was examined by CCK-8 assays (n = 5 per experiment). B CCK-8 assay quantification of CF proliferation (n = 5 per experiment). C RT-qPCR quantification of collagen I and TGF-β₁ mRNA levels (n = 5 per experiment). D Wound repair assays for assessing CF migration ability (n = 5 per experiment). E Immunofluorescence staining of α-SMA expression (green) and the nuclei (DAPI, blue) (n = 5 per group, ≥ 20 fields per group; scale bar = 100 μm). F, Immunofluorescence DHE staining of superoxide anion levels (n = 5 per group). G, SOD and GSH activity (n = 5 per group). H, RT-qPCR quantification of NOX2 and NOX4 mRNA levels (n = 5 per experiment). *P < 0.05 and **P < 0.01 vs. the control group; ^#P < 0.05 and ^##P < 0.01 vs. the Ang II group