Fig. 1

UVRAG^FS inhibits starvation-induced autophagy in vivo. a Representative images (left panels) and quantification (right panels) of GFP-LC3 puncta in indicated tissues from Dox-treated/untreated iUVRAG^FS mice and control mice crossed with GFP-LC3 transgenic mice, following 48 h of starvation. n = 10 tissue sections pooled from three independent experiments. Scale bars, 10 μm. b Western blot analysis of LC3-I/II, p62 levels, Atg12-Atg5 conjugates, and Atg16 levels in the liver from Dox-treated/untreated iUVRAG^FS mice and control mice following 48 h starvation. The densitometric quantification of the LC3-II/LC3-I and the p62/actin ratios under the indicated conditions are shown (right panels). c Representative electron microscopy (EM) images of the liver from Dox-treated/untreated iUVRAG^FS mice in fed or starved conditions for 48 h. Note the accumulation of lipid droplets (LDs) in starved iUVRAG^FS mice on Dox. Insets highlight LDs being surrounded by autophagic membrane structures. Arrowheads (red) denote autophagic vacuoles. The number and size of LD and the number of autophagic vacuoles (AV) in liver from indicated mice were quantified (right panels). n = 10–30 cells pooled from three independent experiments. Scale bars, 1.0 μm. d Co-immunoprecipitation (co-IP) of autophagy-related proteins with UVRAG in 48 h-starved liver from control and Dox-treated/untreated iUVRAG^FS mice. IgG is a negative control. Actin serves as a loading control. IP, immunoprecipitated; WB, western blot; WCL, whole cell lysates. Data in b–d are from one experiment that is representative of three independent experiments. For all quantifications, data (mean ± SD) were from the indicated number of independent experiments and analyzed with two-way ANOVA. n.s., not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****, P < 0.0001. Source data are provided as a Source Data file. See Supplementary Fig. 9 for uncropped data of b, d.