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. 2022 Mar 22;7(6):e153672. doi: 10.1172/jci.insight.153672

Figure 5. Inhibition of PIM1 signaling pathway reduces profibrotic gene expression in IPF-derived lung fibroblasts.

Figure 5

(A) qPCR analysis of profibrotic gene expression in IPF-derived lung fibroblasts transfected with scrambled siRNA or siRNA for NFATc1 for 48 hours, followed by treatment with 2 ng/mL of TGF-β for an additional 24 hours. Data are shown as mean ± SEM of n = 4 independent experiments. P values were calculated using 1-way ANOVA with Holm-Šidák post hoc test. (B) IPF-derived lung fibroblasts were transfected with scrambled or NFATc1 siRNAs for 48 hours and then treated with 2 ng/mL of TGF-β for an additional 24 hours, followed by Western blotting analysis. Shown is a representative blot of 3 independent experiments. (C) qPCR analysis of profibrotic gene expression in IPF-derived lung fibroblasts treated with VIVIT (5 μM) and 2 ng/mL of TGF-β for 24 hours. Data are shown as mean ± SEM of n = 4 independent experiments. P values were calculated using 1-way ANOVA with Holm-Šidák post hoc test. (D) IPF-derived lung fibroblasts were treated with VIVIT (5 μM) and 2 ng/mL of TGF-β for 24 hours and analyzed via Western blot. Shown is a representative blot of n = 2 independent experiments. (E) qPCR analysis of profibrotic genes in IPF-derived lung fibroblasts cotreated with 10 μM of AZD1208 and 2 ng/mL of TGF-β for 24 hours. Data are shown as mean ± SEM of n = 3 independent experiments. P values were calculated using 1-way ANOVA with Holm-Šidák post hoc test. (F) IPF-derived lung fibroblasts were cotreated with 10 μM of AZD1208 and 2 ng/mL of TGF-β for 24 hours and analyzed by Western blotting. Shown is a representative blot of 3 independent experiments. (G) IHC of IPF-derived fibroblasts cotreated with 10 μM of AZD1208 and 2 ng/mL of TGF-β for 24 hours. Scale bar: 50 μm. Representative images of 3 independent experiments are shown.