Figure 2.

miR-1468-3p Promotes Fibrosis, and Targeting miR-1468-3p Reduces TGF-β1-Induced Fibrosis

(A and B) Human cardiac fibroblasts (hCFs) were treated with 20 nM mimic-1468-3p or mimic-Ctrl. After 24 h, cells were fixed and stained for total collagen deposition, or collected for WB analysis. Shown are quantification of total collagen deposition (A) and WB and densitometry analysis for collagen 1, periostin (Postn), and CTGF expression (B). (C–E) hCFs were treated with 50 nM antagomir of miR-1468-3p (antimiR-1468-3p) or control sequence (antimiR-Ctrl). After 24 h, hCFs were treated TGF-β1 (5 ng/mL) where indicated. At the end of the experiment, cells were fixed and stained for total collagen deposition, collected for WB analysis, or collected for qPCR analysis. Shown are quantification of total collagen deposition (C); qPCR analysis for COL1A1, connective tissue growth factor (CTGF), and Postn (D); and WB and densitometry analysis (lower panel) for collagen 1, Postn, and CTGF expression (E). (F) qPCR analysis for miR-1468-3p expression. Vinculin was used as a loading control for WB. Collagen deposition, N = 5–6 per group; qPCR, N = 3–4 per group. Data are expressed as mean ± SD. One-way ANOVA followed by Tukey’s post hoc test and Student’s t test was used. ^$p < 0.05, ^$$p < 0.01 versus mimic-Ctrl; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus antimiR-Ctrl; ^##p < 0.01, ^###p < 0.001 versus antimiR-Ctrl+TGF-β1.