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. 2018 Dec 4;47(4):2011–2028. doi: 10.1093/nar/gky1209

Figure 4.

Figure 4.

RPS3 interacts with SIRT1 mRNA. (A) Heatmap showing genes that were significantly influenced by RPS3. (B) GO enrichment analysis of the RPS3-regulated genes. (C) SIRT1 mRNA was immunoprecipitated by the anti-RPS3 antibody. *P < 0.05, **P < 0.01, and ***P < 0.001. (D–I) Identification of the specific binding region of RPS3 on the SIRT1 mRNA. Different biotinylated fragments were subjected to biotin pull-down assays, followed by western blotting. Input represents 1% of lysate used in pulldown reactions. UN indicate a control pulldown containing beads only. NC indicate negative control pulldown containing no blot-RNA. SIRT1 mRNA and RPS3 interaction was assessed using Western blotting of the RNA-bead complexes and detection with anti-RPS3 antibodies. GAPDH detection was used as a negative control. (J, K) WT 3′UTR-3448-3530 and three mutants M1, M2 and M3 were subjected to RNA pull-down assays, followed by western blotting.