Figure 3. Effect of silencing RPL13 induced a p53-dependent cell cycle arrest.

(A) UACC 903 cells were transfected with siRNA targeting selected RPLs or scrambled control followed by western blotting. Erk2 served as the control for equal protein loading. (B) FF2441 fibroblast and UACC 903, 1205 Lu, or WM35 melanoma cell lines were transfected with RPL13 or scrambled control siRNA to measure increases in p53 and p21 levels by western blotting. Erk2 was used as a control for equal protein loading. (C) UACC 903 and 1205 Lu cells transfected with RPL13 or scrambled control siRNA were treated with cycloheximide (CHX) to stop protein synthesis. Western blotting from 0–2 hours after CHX treatment showed stability of p53. The siScramble 0–2hr blots were intentionally overexposed to enable comparison to the other blots. Enolase served as the control for equal protein loading. (D) UACC 903 or 1205 Lu cells were transfected with RPL13 siRNA alone or in combination with p53 siRNA and after 3 days incubation, cells were stained with propidium iodide, run on a BD FACSCalibur and results analyzed by ModFit LT (N≥3).