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. 2019 Feb 5;38(5):e99599. doi: 10.15252/embj.201899599

Figure 3. EZH2 binds with cap‐independent translation complex and increases p53 protein translation.

Figure 3

  1. List of top hits of EZH2‐bound proteins identified by TAP‐MS. The number of peptides and coverage of each protein are indicated.
  2. Reciprocal co‐IP of endogenous EZH2 and eIF4G2 or PABP1 in C4‐2 cells.
  3. Co‐IP of endogenous EZH2 with eIF4G2 or PABP1 from C4‐2 cell lysate pre‐treated with RNaseA in 37°C for 30 min. The effectiveness of RNaseA treatment was monitored by RT–PCR analysis of the presence of GAPDH mRNA.
  4. C4‐2 cells were transfected control (siC) or EZH2‐specific siRNA and lysed for polysome fractionation. RNA was extracted from even‐number fractions followed by RT–qPCR analysis for p53 and GAPDH mRNA. The β‐ACTIN mRNA was used as an internal control. Data shown as means ± SD (n = 3). Statistical significance was determined by two‐tailed Student's t‐test. *P < 0.01; NS, no significance.
  5. C4‐2 cells were transfected with control (siC) or EZH2‐specific siRNAs in combination with empty vector, EZH2 WT, or different deletion mutants followed by polysome fractionation and RT–qPCR. β‐ACTIN mRNA was used as an internal control. Data shown as means ± SD (n = 3). *P < 0.01.

Source data are available online for this figure.