Fig. 6

LanCL1 restores impaired AKT activity and mitigates oxidative stress in ALS mice. Immunoblotting (a) and quantifications (b) of pAKT (S473), pAKT (T308), and BAX in the lumbar spinal cord (L3–L5) of indicated mice. Data represent mean ± SEM; WT n = 4 mice, LanCL1 cKI n = 3 mice, G93A n = 6 mice, G93A; LanCL1 cKI n = 4 mice; *p < 0.05, **p < 0.01, ***p < 0.001; n.s. nonsignificant; one-way ANOVA followed by Tukey’s post hoc test. Immunoblotting (c) and quantifications (d) of HO-1 in the lumbar spinal cord (L3–L5) of indicated mice. Data represent mean ± SEM, n = 4 mice per genotype, *p < 0.05, **p < 0.01, one-way ANOVA followed by Tukey’s post hoc test. e HeLa cells were transfected with 2 μg of Myc-pRK5 vector or Myc-tagged LanCL1. Twenty four hours later cells were pretreated with either dimethyl sulfoxide (DMSO), BKM120 (5 μM), or LY294002 (20 μM) for 30 min prior to addition of 150 μM H₂O₂. Apoptosis assay was performed by Hoechst 33342 staining 24 h after the H₂O₂ treatment. f The percentage of apoptotic HeLa cells with fragmented or condensed nuclei was determined by Hoechst 33342 staining. At least 1000 cells per condition were scored for Hoechst 33342 staining. Data represent mean ± SEM of four independent experiments, **p < 0.01, ***p < 0.001, two-way ANOVA followed by Bonferroni post hoc test. g Immunoblotting showing that the PI3K inhibitors (BKM120 and LY294002) effectively blocked AKT activation in response to H₂O₂ treatment in HeLa cells. h Diagram illustrating that LanCL1 regulates cell survival through mitigating oxidative stress and enhancing AKT phosphorylation