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. 2013 Oct 15;27(20):2246–2258. doi: 10.1101/gad.221739.113

Figure 4.

Figure 4.

The 3′ UTR, but not the 5′ UTR, of p53 mRNA is required for S195D to enhance p53 expression. (A–C) H1299 cells were transiently cotransfected with a control pcDNA3 vector or various amounts of pcDNA3 vector expressing HA-tagged wild-type RNPC1 (A), S195D (B), or S195A (C) along with a fixed amount of p53 expression vector that contains the coding region (ORF) alone or in combination with the 5′ UTR, 3′ UTR, or both. Twenty-four hours post-transfection, cell lysates were collected, and the levels of p53, RNPC1, and actin were analyzed by Western blot analysis. The level of p53 protein was normalized to that of actin, and the relative fold change is shown below each lane. (D–G) RNPC1−/−; p53−/− MEFs were transiently cotransfected with a p53 expression vector that contains the coding region (ORF) alone (D) or together with the 5′ UTR (E), 3′ UTR (F), or both (G) along with a control vector or a pcDNA3 vector expressing HA-tagged wild-type RNPC1, S195D, or S195A for 24 h. Cell lysates were collected and subjected to Western blot analysis to determine the level of p53, RNPC1, and actin. The level of p53 protein was normalized to that of actin, and the relative fold change is shown below each lane.