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. 2020 Jul 23;5(14):e137127. doi: 10.1172/jci.insight.137127

Figure 5. p300 mediates Nox4 gene expression in lung fibroblasts.

Figure 5

(A–C) Primary IPF fibroblasts were transfected with nontargeting (NT) siRNA or p300 siRNA. (A) Representative Western blots of fibroblasts from 2 different individuals with IPF transfected with siRNA NT or p300 demonstrated the levels of p300 and β-actin. (B) The densitometry of p300-associated signals detected (ratio to β-actin) in A. *P < 0.05, compared with NT control of the same cell line, by 2-tailed t test. (C) The expression of Nox4 mRNA was analyzed by real-time quantitative PCR of the IPF cells transfected with siRNA NT or p300 (mean ± SD, n = 3 experimental replicates of a representative cell line). *P < 0.05 compared with NT control, by 2-tailed t test. (D–F) IMR90 cells were transfected with siRNA NT or p300, and then the cells were treated with TGF-β1 (2 ng/mL) for 48 hours and analyzed for p300 expression by Western blot and Nox4 mRNA expression by real-time PCR. (D) Representative Western blots of p300 and β-actin. (E) Densitometric analysis of p300-associated signals detected (ratio to β-actin) in D. *P < 0.05, TGF-β1–treated cells with p300 siRNA vs. NT siRNA, by 2-tailed t test. (F) The mRNA expression of Nox4 by real-time PCR (mean ± SD, n = 3 experimental replicates). *P < 0.05, compared with vehicle control; #P < 0.05, the transfected cells with TGF-β1 treatment, siRNA p300 vs. NT, by 2-tailed t test.