Figure 6

MiR-145-3p and miR-891a-3p control the fibrotic response while miR-223-3p, miR-486-3p, and miR-488-5p control the inflammatory response. Quantification of mRNA levels of (A) hypertrophic (ANF, BNP, and αSKA), (B) fibrotic (CTGF, COL1α1, and α-SMA), and (C) inflammatory markers (TNFα, ICAM and MR) in respectively nRCMs, nRCFs, or BMDMs after miRNA mimic transfection (Unpaired t-test). (D) Representative western blots of collagen type I (Col I) and normalizer α-Tubulin (α-Tub) performed with cell lysates of nRCFs transfected with negative control (NEG), miR-145-3p, miR-151a-5p, and miR-891a-3p mimics. Samples were run on the same gel and blot, different regions of the blot were cropped to quantify Col I and a-Tub, and every sample per protein was acquired under the same exposure conditions. (E) Western blot quantification reveals increased and decreased Collagen Type I protein expression in miR-145-3p and miR-891a-3p transfected nRCFs, respectively (One Way ANOVA). (F) Levels of cytokines and chemokines determined using a Mouse Cytokine Array in the supernatant of BMDMs transfected with miR-223-3p, miR-486-3p, or miR-488-5p mimics. In (A–C), bar graphs display mean log₂ fold change in comparison with negative control transfected cells ± standard error. In (E,F), bar graphs display mean fold change in comparison with negative control transfected cells ± standard error. *Denotes P < 0.05, **denotes P < 0.01, and ***denotes P < 0.001 versus unstimulated negative control.