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. 2020 Jun 2;39(13):e103838. doi: 10.15252/embj.2019103838

Figure 2. The IRBC and ATR distinctly regulate p53 stabilization upon nucleotide depletion.

Figure 2

  1. HCT116 cells were transfected with either a NT or a siRNA against RPL11 for 24 h and treated with the vehicle alone (−) or the indicated concentration of MPA for 24 h. The levels of p53 and p21 were analyzed on Western blots. GAPDH was used as a loading control. Quantification of band intensity of p53 and p21 of four independent experiments is shown (right panel).
  2. HCT116 cells were transfected with the indicated siRNA and 24 h later treated with the vehicle alone or 10 μM MPA for 6 h, and the levels of p53 and p21 were analyzed on Western blots. GAPDH was used as a loading control. Quantification of band intensity of p53 of at least four independent experiments is shown (right panel).
  3. HCT116 cells were transfected with either a NT siRNA or a siRNA against RPL11 for 24 h and were preincubated for 30 min in the absence (−) or the presence (+) of the combination of 10 μM of the ATR inhibitor VE‐821 (ATRi) and 10 μM the ATM inhibitor KU‐55933 (ATMi), followed by addition of the vehicle alone (−) or MPA 10 μM for additional 24 h. The levels of p53 were analyzed on Western blots. GAPDH was used as a loading control. Quantification of band intensity of p53 of at least two independent experiments is shown (right panel).
Data information: All data are presented as Mean ± SD, relative to control. *P < 0.05, **P < 0.01, ***P < 0.001, by two‐tailed Student's t‐test.