Fig. 3: TTYH1 mitigates the inhibitory effect of C1P on autophagic flux. | Nature Communications

Fig. 3: TTYH1 mitigates the inhibitory effect of C1P on autophagic flux.

From: Endolysosomal processing of neuron-derived signaling lipids regulates autophagy and lipid droplet degradation in astrocytes

Fig. 3

a Schematic representation of TTYH1-mediated endolysosomal clearance of C1P. b Ttyh1 deficiency exacerbates autophagic flux inhibition by exogenous C1P. Levels of p62 and LC3 in cKO astrocytes in response to exogenous C1P were analyzed. Astrocytes were treated with 0 (0.1% DMSO), 0.1, or 0.5 µM of C1P in 0.1% BSA-supplemented culture media for 2 h. Shown are representative immunoblot images and quantifications of three independent experiments. Values were normalized to those of Ttyh1 WT, 0 µM C1P condition on the same blot. Data were presented as mean ± SEM. Number of biological replicates (n) is shown in brackets at the bottom of each bar. ***P < 0.001, **P < 0.01, One-way ANOVA (p62/αTub: F = 56.91, df =  17; LC3-II/I: F = 42.95, df = 23) followed by Bonferroni’s multiple comparisons post hoc test. c Human TTYH1 rescues C1P-induced autophagic flux blockage in Ttyh1 KO astrocytes. Levels of LC3 and p62 in Ttyh1 KO astrocytes in response to C1P (0.5 µM) were analyzed. Shown are representative immunoblot images and quantifications of four independent experiments. Values were normalized to those of the untreated mock-transfected control on the same blot. Data are presented as mean ± SEM. Number of biological replicates (n) is shown in brackets at the bottom of each bar. One-way ANOVA (p62/αTub: F = 15.43, df = 23; LC3-II/I: F = 40.88, df = 23) followed by Bonferroni’s multiple comparisons post hoc test. n.s. not significant, P > 0.9999. d Overexpression of TTYH1 in astrocytes restores autophagic flux upon C1P. Levels of LC3 and p62 in primary human brain astrocytes in response to C1P (50 µM) were analyzed. Shown are representative immunoblot images and quantifications of four independent experiments. Values were normalized to those of the untreated mock-transfected control on the same blot. Data were presented as mean ± SEM. Number of biological replicates (n) is shown in brackets at the bottom of each bar. One-way ANOVA (p62/αTub: F = 17.98, df = 23; LC3-II/I: F = 117.5, df = 23) followed by Bonferroni’s multiple comparisons post hoc test. n.s. not significant, P > 0.9999. Source data are provided as a Source Data file.

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