Fig. 7
From: A truncating mutation in the autophagy gene UVRAG drives inflammation and tumorigenesis in mice
UVRAGFS activates β-catenin by promoting age-related autophagy suppression. a H&E and IHC of colon from Dox-treated control and iUVRAGFS mice (12-month-old). Data are from one animal that is representative of 5–12 animals in each group. b Crypt length of the colon in a. c, d Representative images (c) of Ki67, cleaved caspase 3, and γ-H2AX of colons in 12-month-old control and iUVRAGFS mice on Dox and quantitation (d) of Ki67- and γ-H2AX-positive cells per crypt of indicated genotype. Arrow indicates apoptosis. e, f IHC of β-catenin and c-Myc (e) and quantitation of nuclear β-catenin per crypt (f) from control and iUVRAGFS mice (12-month-old). g WB of indicated protein expression in colons of the indicated genotype. h Quantitative RT-PCR of β-catenin and its target gene expression in colons from 18-month-old mice. n = 3. i, j WB of β-catenin and autophagy marker proteins in iFHC cells treated with 3-MA (1 mM) (i) or with chloroquine (CQ, 20 μM). k UVRAGFS inhibits β-catenin interaction with autophagy proteins. SW480 cells were transfected with increasing amounts of Flag-UVRAGFS. WCL were used for IP with anti-β-catenin, followed by IB with the indicated antibodies. The densitometric quantification of the p62/actin ratio is shown (bottom). l Quantitative RT-PCR of indicated gene expression in cells in k. n = 3. m Cell cycle analyses of SW480 cells stably expressing vector or UVRAGFS. n = 3. n Co-IP of β-catenin with LC3 and TCF4 in spleens from 18-month-old mice. Data in b–f are from 10 to 20 HPF pooled from three independent experiments. Data in g, n are from two randomly chosen samples per group with similar results observed in all ten samples per genotype. For all quantifications, data (mean ± SD) were analyzed with Student’s t-test (b, d, f), two-way ANOVA (h, m) or one-way ANOVA (k, l, m). n.s., not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Scale bars, 100 μm. Source data are provided as a Source Data file. See Supplementary Fig. 9 for uncropped data of g, i, j, k, n and Supplementary Fig. 10 for raw data of m.
