Fig. 5

RBMS3 destabilized Twist1 transcript by directly binding to the 3′-UTR of Twist1 mRNA. a, b in SUM-1315 and MDA-MB-231 cell lines, RBMS3 overexpression shortened the half-life of Twist1 mRNA, while knockdown of RBMS3 prolonged the halflife of Twist1 mRNA. Overexpression (RBMS3) and control cells (NC), knockdown (shRBMS3) and the control (SCR) were treated with Act D at a concentration of 5 μg/ml. The total RNA were extracted at 0, 1, 2, 4, 6, and 8 h, respectively, and then followed by qRT-PCR analysis. d, e SUM-1315 and MDA-MB-231 cells lysates were immunoprecipitated with RBMS3 or IgG antibody and analyzed by using RT-PCR and qRT-PCR to detect Twist1 and Smad3 transcript levels. c Schematic diagram of various regions in the 3′-UTR of Twist1 mRNA. f, g The reporter containing Twist1 3′-UTR-B, −C was decreased by overexpression of RBMS3 in SUM-1315 and MDA-MB-231 cells. Data were shown as mean ± SEM, *P < 0.05