Fig. 5

Knockout of Rubicon ameliorates age-associated phenotypes in mice. a Rubicon protein level in kidney was elevated in 20-month-old mice relative to 2-month-old mice. Western Blot from WT mice and Rubicon KO (KO) mice shows the specific bands of Rubicon. b A quantification of Western Blot shown in a. Value represent means ± s.e.m (2 months, n = 3; 20 months, n = 3). P value (*P < 0.05) was determined by t-test. c Western blotting samples showing LC-3 and p62 in mouse kidney at 20 months of age. d Immunohistochemical images of 20-month-old kidney showing that collagen I–positive area was reduced in Rubicon-knockout (Rubicon KO) kidney relative to WT. P value (*P < 0.05) was determined by t-test. e A quantification of collagen I–positive area in d. Values represent means ± s.e.m (WT, n = 5; Rubicon KO, n = 6). P value (*P < 0.05) was determined by t-test. f qRT-PCR analysis of several fibrosis markers (TGF-b1, Col1a1) in 20-month-old kidney. Values represent means ± s.e.m (WT, n = 5; Rubicon KO, n = 6). P value (*P < 0.05) was determined by t-test. g Lewy body and Lewy neurite–like inclusions containing phosphorylated α-Syn (green punctae, indicated by open arrowheads) were less abundant in neuronal Rubicon-knockout (Rubicon^flox/flox: Nestin-Cre) mice than in controls (Rubicon^flox/+: Nestin-Cre), 10 months after the injection. h Percentages of mice having small (<100), moderate (100–200), or large (>200) numbers of phosphorylated α-Syn positive signals. P-value (*P < 0.05) was determined by χ² test (Rubicon^flox/+: Nestin-Cre, n = 7; Rubicon^flox/flox: Nestin-Cre, n = 6). Scale bars, 100 μm (d); 500 μm (g)