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. 2021 Jan 1;11(6):2706–2721. doi: 10.7150/thno.48679

Figure 4.

Figure 4

Inhibition of JMJD3 by GSKJ4 or siRNA promotes activation of renal interstitial fibroblasts in culture exposed to TGFβ1. (A, D) NRK-49F cells were incubated with medium containing 0.5% serum or treated with TGFβ1 (2 ng/mL) in the presence or absence of GSKJ4 (6 μM) for 36 h. Cell lysates were prepared and subject to immunoblot analysis with antibodies against JMJD3, H3K27me3, β-actin (A), α-SMA, fibronectin, collagen III, β-tubulin (D). Expression levels of JMJD3 (B), H3K27me3 (C), α-SMA (E), fibronectin (F), collagen III (G), were quantified by densitometry analysis and then normalized with β-tubulin or β-actin, respectively. (H, K) NRK-49F cells were transfected control siRNA or JMJD3 siRNA and then incubated with medium containing 0.5% serum or treated with TGFβ1 (2 ng/mL) for 36 h. Cell lysates were prepared and subject to immunoblot analysis with antibodies against JMJD3, H3K27me3, β-actin (H) α-SMA, fibronectin, collagen III, β-tubulin (K). Expression levels of JMJD3 (I), H3K27me3 (J), α-SMA (L), fibronectin (M), and collagen III (N) were quantified by densitometry analysis and then normalized with β-tubulin or β-actin, respectively, as indicated. Values are the means ± sem of at ≥3 independent experiments. *P < 0.05; **P < 0.01.