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. Author manuscript; available in PMC: 2015 Jun 15.
Published in final edited form as: Cancer Res. 2014 Apr 24;74(12):3218–3227. doi: 10.1158/0008-5472.CAN-13-1706

Figure 3. MiR-542-3p induces ribosomal p53 response.

Figure 3

(A) MiR-542-3p induces ribosome abnormality. U2OS cells were transfected as indicated. Forty-eight hours later, RNA was extracted and run on Agilent Bioanalyser. The 28S/18S ratio was calculated from electropherogram data. (B) MiR-542-3p suppresses the expression of RPS23, RPS28 and RPL22. U2OS and HCT116 cells transfected with negative or miR-542-3p mimics were analyzed for the expression of ribosome subunits by western blotting. (C) Effect of RPL22, RPS23 and RPS28 depletion by siRNA on 18S rRNA, RPL11 and p53 levels. U2OS cells transfected as indicated were collected for RNA quality analysis by Agilent Bioanlyser (left panel) or western blot analysis (right panel). (D) Depletion of RPS23 decreases MDM2-p53 interaction and p53 polyubiquitination. U2OS cells transfected as indicated were treated with MG132 (2μM) for 4hr and then lyzed for immunoprecipitation with anti-p53 followed by western blot analysis. (E) miR-542-3p and siRPS23 increase RPL11 and its association with MDM2. Cells transfected as indicated were lyzed for immunoprecipitation with anti-MDM2 followed by western blot analysis. (F) Depletion of siRPL11 reverses miR-542-3p and siRPS23-mediated p53 induction. U2OS cells were co-transfected as indicated were lyzed for western blot analysis. *, P < 0.01.