RUNX2 does not significantly regulate the expression levels of either hsa-mir-30c or NOV. (A, B) Detection of NOV and hsa-mir-30c levels in MDA-MB-231 stably expressing empty vector (EV), wild-type Runx2 (WT), or R398A/Y428A mutant Runx2 (RY). (A) Representative Western blots of MDA-MB-231 stable cell lysates. Top blot: RUNX2 (top band: transgenic murine Runx2, lower band: endogenous human RUNX2). Middle blot: NOV. Lower blot: Lamin C. (B) qPCR detection for hsa-mir-30c in consecutive (N = 2) passages of stable MDA-MB-231 cells normalized to U6 snRNA. (C, D) Detection of NOV and hsa-mir-30c following 48 h of transient transfection of non-targeting siRNA (NS) and RUNX2 siRNA (siR2). (C) Representative Western blots of MDA-MB-231 lysates following 48 h of siRNA transfection. Vertical dashed line indicates that the image of the blot was cut for figure. Top blot: RUNX2. Middle blot: NOV. Lower Blot: α-Tubulin. (D) qPCR detection of hsa-mir-30c levels of two technical replicates each of four biological replicates following 48 h transfection of siRNA. p-value from Student’s t-test with Welch correction: approaching statistical significance (P-value = 0.0835). (B, D) Bars equal mean, error bars equal SEM.