Ctrl IR, 2h G2 P53 Ser15 S G1 G2 S G1 Figure S1. The bivariate (phospho-p53 Ser15 vs DNA content) distribution of control and irradiated with 6 Gy mESCs.

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Ctrl IR, 2h G2 P53 Ser15 S G1 G2 S G1 Figure S1. The bivariate (phospho-p53 Ser15 vs DNA content) distribution of control and irradiated with 6 Gy mESCs. Measurements were carried out 2 h post-irradiation. G1/S S G2/M Control 16.2% 60,6% 23,2% + pifithrin-α 16,5% 58,2% 25,3% + NaBut 28,6% 55,1% 16,3% 30,4% 48,5% 21,1% Figure S2. Cell cycle distribution of control, treated with pifithrin-α (10µM), treated with NaBut (4 mM), treated with NaBut + pifithrin-α. Measurements were carried out 24 h post treatment.

A Ctrl IR, 1d G1: 26% S: 56% G2/M: 18% G1: 31% S: 47% G2/M: 22% B C relative cell viability, % Relative fluorescence units Ctrl 1d 3d 1d 3d 1d, 6 Gy 6 Gy 6 Gy 3 Gy 3 Gy +ZVAD Ctrl 1d 1d 3 Gy 6 Gy D E Relative fluorescence units relative cell viability, % Ctrl Nutlin, 1d Nutlin,1d +ZVAD Ctrl Nutlin, 1d Figure S3. Irradiation and nutlin treatment caused a significant level of apoptosis in mESCs. (A) Cell cycle distribution of control and irradiated (6Gy, 24h) mESCs. (B): In vitro caspase-3 activity assay of mESCs irradiated with 3 Gy and 6 Gy and examined 1 and 3 days post-irradiation. Data are presented as mean ±S.E.M. of three independent replicates (n=3). Caspase inhibitor ZVAD was added to the reaction sample 1 day after IR 6Gy to use as a negative control. (C): MTT assay. The effect of irradiation by a dose of 6 Gy on mESCs viability. Measurements were carried out 24 h post-irradiation. (D) In vitro caspase 3 activity assay of control mESCs and treated with nutlin (10 µM) for 1 day. A sample with caspase inhibitor ZVAD added to the reaction served as a negative control. Data are presented as mean ±S.E.M. of three independent replicates (n=3). (E) MTT assay. The effect of nutlin treatment (10 µM) on mESCs viability.