Extended Data Fig. 1: Low-protein (LP) diets aggravate intestinal DNA damage and inflammation.

a, Representative IHC images of colon tissues from mice fed with AIN-93G, 6% protein, or 60% protein (high-protein, HP) diets and water containing 2% DSS. b, Quantification of IHC of γH2A.X in crypts of colon tissues from mice fed with AIN-93G or 60% protein diets and water containing 2% DSS. n = 73 (AIN-93G) and 69 (60% protein) crypts. 3 independent experiments. c, Quantification of IHC of inflammatory cells markers in colon tissues from mice in (b). Sample size (n) see SourceData_Extended_Fig1. d, Representative IHC images of p-ATR and γH2A.X in colon tissues from mice fed with AIN-93G or LP diets. DDR was induced by intraperitoneal injection of 5-FU. n = 3 mice. e, Western blot of p-CHK1 and γH2A.X in HeLa cells in response to HU treatment and deprivation of individual amino acid. n = 3 independent experiments. f, Relative quantification of p-CHK1 in HeLa cells in response to HU treatment and deprivation of individual amino acid. Normal culture medium as control. n = 3 independent experiments. g, Relative quantification of γH2A.X level in HeLa cells in response to HU treatment and deprivation of individual amino acid. Normal culture medium as control. n = 3 independent experiments. h, Western blot of p-ATR, p-CHK1 and γH2A.X in HeLa cells. 2 mM HU was added to control or GCN2 knockout HeLa cells with or without amino acids starvation. n = 3 independent experiments. i, j, Knockout of GCN1 (i) (n = 3 independent experiments.) or DEPDC5 (j) (n = 3 independent experiments) does not rescue the levels of DDR markers downregulated by amino acids deprivation in HeLa cells. Western blot analyses of levels of p-ATR, p-CHK1 and γH2A.X in cells in response to HU treatment with or without amino acids starvation. Scale bar, 50 µm. Error bars represented mean ± s.d. Statistical comparisons were made using Mann–Whitney U-test (b, c) or one-way ANOVA test (f, g, h, j). NS, not significant.

Source data