Fig. 4: Asparagine maintains cell survival, and represses p53-dependent cell cycle checkpoint activation. | Nature Communications

Fig. 4: Asparagine maintains cell survival, and represses p53-dependent cell cycle checkpoint activation.

From: p53-mediated control of aspartate-asparagine homeostasis dictates LKB1 activity and modulates cell survival

Fig. 4

a HCT116 cells transfected with control or p53 siRNA were cultured with 0, 1 or 2 U per ml ASNase for 48 h. Cell survival (% of viable cells) was determined by trypan blue exclusion assay. b Survival of and Protein expression in MEFs transfected with control, p53 siRNA and/or ASNS siRNA as indicated. c, d Survival of and protein expression in HCT116 cells (c) and MEFs (d) transfected with control, ASNS siRNA and/or p21 siRNA as indicated. e Protein levels of p53, p21 and phosphorylated p53 (S15) in p53+/+ and p53−/− HCT116 cells transfected with control or ASNS siRNA for 3 days. f Protein expression in p53+/+ and p53−/− HCT116 cells treated with 0, 1 or 2 U per ml ASNase for 48 h was analysed. g Protein expression in siCtrl and siASNS HCT116 cells cultured in medium containing increasing amounts of Asn for 3 days was analysed. h Cell cycle distribution of siCtrl and sip53 HCT116 cells treated with 0 or 2 U/ml ASNase for 48 h. The percentages of cells in the sub-G1, G1, S and G2 phases were determined by PI staining and flow cytometry analysis. i p53+/+ and p53−/− HCT116 cells transfected with control or ASNS siRNA were cultured in medium containing 0 or 0.1 mM Asn for 3 days. Cell cycle distribution was determined. j Cell cycle distribution of HCT116 cells transfected with control, ASNS siRNA and/or p21 siRNA for 72 h. Data are mean ± s.d., unpaired two-tailed Student’s t-test, *p < 0.05, **p < 0.01, ***p < 0.001, NS not significant. Source data are provided as a Source Data file.

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