Fig. 2: Neuronal signalling controls ubiquitin-dependent turnover and PolyQ aggregation. | Nature Communications

Fig. 2: Neuronal signalling controls ubiquitin-dependent turnover and PolyQ aggregation.

From: Thermosensation in Caenorhabditis elegans is linked to ubiquitin-dependent protein turnover via insulin and calcineurin signalling

Fig. 2

a Schematic illustration of analysed C. elegans ciliated head and tail neurons. b WT, dyf-1(mn335), or worms harbouring mutations affecting the functionality of indicated neurons and expressing UbV-GFP were analysed by Western blotting against GFP and tubulin. Mutants: AWA odr-7(tm4791), AWB/ADF lim4(ky403), AWC ceh-36(ks86), ASI unc-3(e151), ASH unc-42(e270), ASE che-1(ot75), PQR, PHA, PHB ceh-14(ch3), ASG/ADL lin-11(n389), ASK ttx-3(ot22), and IL2 unc-86 (n847). c Quantification of worms from (b), n = 6 (WT, dyf-1, ASH, ASE), 5 (ASI, ASK), 4 (AWA, AWB/ADF, PQR, PHA, PHB, ASG/ADL) and 3 (IL2, AWC) biologically independent experiments. Column graphs: mean ± standard deviation, scatter plots: individual experiments, dyf-1(mn335) p = <0.0001, odr-7(tm4791) p = 0.001 (including dyf-1(mn335) for maximum changes), lim4(ky403) p = 0.5413, ceh-36(ks86) p = 0.9996, unc-3(e151) p = 0.9996, unc-42(e270) p = 0.0067, che-1(ot75) p = 0.6447, ceh-14(ch3) p = 0.0528, lin-11(n389) p = 0.708, ttx-3(ot22) p = 0.4099, unc-86(n847) p = 0.9867 (excluding dyf-1(mn335) from comparison to detect significant changes compared with WT), One-way ANOVA with Dunnett’s multiple comparisons test. d, e Indicated mutant worms analysed as in (b), n = 2 biologically independent experiments (d, e). f Fluorescence and Nomarski images of day 1 WT or dyf-1(mn335) worms expressing intestinal Q44::YFP. Scale bars: 200 μm. g Quantification of Q44::YFP aggregates per day 1 worm. Red: mean ± s.e.m. WT versus: dyf-1(mn335) p < 0.0001, gcy-8 (oy44) p = 0.0072, dyf-1(mn335) versus gcy-8(oy44) p = 0.4192, Kruskal–Wallis one-way ANOVA with Dunn’s multiple comparisons test, n = 2 biologically independent experiments (c, g) ****p ≤ 0.0001, ***p ≤ 0.001, **p ≤ 0.01, n.s. p > 0.05. h Confocal images of the finger-shaped amphid AFD (Pgcy-8::GFP, villi and cilium) and branched AWA (ODR-10::GFP) cilia in WT and indicated mutants. Anterior: left. Arrowheads: accumulation of ODR-10::GFP in the cilia. Typical results of (Pgcy-8::GFP: 27 (WT), 30 (dyf-1(mn335)); odr-10::GFP: 45 (WT), 29 dyf-1(mn335), 37 dyf-1(mn335); gcy −8(oy44) worms, n = 2 biologically independent experiments. Scale bar 5 μm. See also Supplementary Fig. 2f, Source data are provided as a Source Data file.

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