Mircury lnatm array

MicroRNA array analysis was done by using miRCURY LNA^TM Array microRNA profiling services (Exiqon, Vedbaek, Denmark). The sample quality of total RNAs from three samples of both the controls and the stretched samples at each time point was assessed using Bioanalyser 2100 (Agilent), and those RNA integrity number (RIN) values that were greater than 7.6. Samples (n = 3 biological replicates in all timepoints in all groups except n = 2 at 1-hour time point) were labeled with miRCURY LNA^TM Hy3^TM/Hy5^TM Power labeling kit and hybridized on the miRCURY LNA^TM Array (v.10.0) probes based on miRBase 11.0 annotation. Quantified signals were normalized using the global Locally WEighted Scatterplot Smoothing (LOWESS) regression algorithm. MicroRNAs were defined as differentially expressed by using two-tailed t-test (P < 0.05) for comparing experimental groups of I) 1, 4 and 12 hours, and II) 24 and 48 hours of stretch against their own control.

Co-cultured or control ASMC-derived total RNA was analyzed for miRNA expression by miRCURY LNA^TM Array microRNA seventh generation profiling services (Exiqon). P value correction utilizing Benjamini–Hochberg false discovery rate was applied, and data analysis was conducted by Exiqon.