Figure 2.

miR-148a Targets gp130 and Differentially Regulates gp130 Downstream Signaling in Concentric and Eccentric Cardiac Remodeling

(A) Location and evolutionary conservation of the mmu-miR-148a seed region on gp130 3′ UTR (Ensemble: ENSMUST00000183663.7). (B) Schematic representation of luciferase reporter constructs. (C) Activity assay of luciferase reporter constructs in Cos7 cells. (D) Western blot for endogenous gp130 in neonatal rat cardiomyocytes when transfected with scrambled control antimir (scr.anti-miR), antimir-148a (anti-miR-148a), or precursor for miR-148a (pre-miR-148a) and quantification of tubulin-corrected gp130 western blot signals. (E) Design of the study: 2-month-old mice were subjected to sham or TAC surgery for 2, 4, 6, or 8 weeks. Mice were subjected to echocardiographic evaluation and hearts harvested at indicated time points. (F) RT-PCR analysis of miR-148a-3p expression at indicated time points. (G–I) Echocardiographically determined LV mass divided by BW (G), EF (H), or LVIDs (I), respectively. (J) Western blot analysis of myocardial CT-1 expression and quantification at indicated time points. (K) Western blot analysis of myocardial gp130 expression and quantification at indicated time points. (L) Western blot analysis of myocardial phosphorylated and unphosphorylated forms of STAT3 and quantification at indicated time points. Data are means ± SEM. One-way ANOVA with Bonferroni’s or Newman-Keul’s multiple-comparison test was used to compare groups. n, number of transfection experiments (C), number of transfected wells (D), number of mice (F–L). TAC, transverse aortic constriction; LV mass divided by BW, left ventricular mass to body weight ratio; EF, ejection fraction; LVIDs, left ventricular internal diameter in systole. *p < 0.05 versus corresponding control group.