Fig. 1. Loss of liver p53 exacerbates liver damage and increases ROS during CCl4-mediated liver regeneration.
Representative H&E images (A) and quantification (B) of damaged area (% per field) in livers from Albumin-Cre; p53WT/WT mice (WT) and Albumin-Cre; p53FL/FL (FL) mice at indicated times (hours) following CCl4 treatment. Damaged areas outlined. Scale bars 100 μm. Higher magnification inset images highlight centrilobular liver damage including vacuolisation (black arrows). These images are taken from different H&E slides than those depicted for damaged area. Scale bars 10 μm. Images reproduced without annotation and at full size in Fig. S1 C/D. Quantifications from N = 2 untreated (0 h) mice/group, N = 6 mice/group at 24 h, N = 5 WT and N = 4 FL mice at 48 h, N = 7 mice/group at 72 h, N = 3 mice/group at 168 h. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p values. ****p < 0.0001. Plasma ALT (C) and AST (D) activity (mU/mL) in Albumin-Cre; p53WT/WT (WT) and Albumin-Cre; p53FL/FL (FL) mice treated as in (A). N = 3 untreated (0 h) mice/group. N = 4 mice/group/time point thereafter. Each data point represents the mean from technical duplicates per mouse. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p-values. *p < 0.05, **p < 0.01, ****p < 0.0001. Staining (E) and quantification of stain area (F) of frozen sections for oil-red-O (ORO) in Albumin-Cre; p53WT/WT (WT) and Albumin-Cre; p53FL/FL (FL) mice treated as in (A). Scale bars 20 μm. Quantification from N = 5 untreated (0 h), 24 h, and 48 h mice/group, N = 7 mice/group at 72 h, and N = 3 mice/group at 168 h. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p values. *p < 0.05, ***p < 0.001. Data from p53FL/FL mice also used in (L) (normal water) but different representative images are shown. IHC staining (G) and quantification (H) of malondialdehyde (MDA) in Albumin-Cre; p53WT/WT (WT) and Albumin-Cre; p53FL/FL (FL) mice at indicated times (hours) after CCl4 treatment. Scale bars 20 μm. Quantification from N = 6 untreated (0 h) mice/group, N = 7 WT and N = 6 FL mice at 24 h, N = 7 mice/group at 48 and 72 h, and N = 3 mice/group at 168 h. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p values. **p < 0.01, ****p < 0.0001. Data from p53FL/FL mice also used in (J) (normal water) but different representative images are shown. IHC staining (I) and quantification (J) of malondialdehyde (MDA) in Albumin-Cre; p53FL/FL (FL) mice given control (water) or 30 mM N-Acetylcysteine-supplemented drinking water (NAC) for one week prior to CCl4 treatment. Images from indicated times (hours) after CCl4 treatment. Scale bars 20 μm. Quantification from N = 6 water and N = 2 NAC untreated mice (0 h), N = 6 water and N = 4 NAC mice at 24 h, N = 7 water and N = 4 NAC mice at 48-72 h. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p-values. **p < 0.01, ****p < 0.0001. Data from p53FL/FL (normal water) mice also used in (H) but different representative images are shown. Staining (K) and quantification (L) of oil-red-O (ORO) in Albumin-Cre; p53FL/FL (FL) mice given control (water) or 30 mM N-Acetylcysteine-supplemented drinking water (NAC) for one week prior to CCl4 treatment. Images from indicated times (hours) after CCl4 treatment. Scale bars 20μm. Quantification from N = 5 untreated water (0 h) and N = 2 untreated NAC mice, N = 5 water and N = 4 NAC mice at 24 and 48 h, N = 7 water and N = 4 NAC mice at 72 h. Data presented as mean ± SEM and analysed using two-way ANOVA with Holm-Sidak’s multiple comparisons test and multiplicity-adjusted p-values. *p < 0.05. Data from p53FL/FL (normal water) mice also used in (F) but different representative images are shown.