Fig. 5: Neuronal signalling primes the intestine to control UPS activity via calcineurin signalling.

a Part of the calcium signalling pathway Cel04020 (KEGG database) with gene expression changes in dyf-1(mn335) compared with WT mapped on it. Pathway nodes correspond to genes coloured in red when gene expression is significantly increased and blue when decreased in dyf-1(mn335), non-affected nodes are coloured in grey, white nodes correspond to genes which were not measured in the microarray experiment. CaN: tax-6 and cnb-1. b Protein lysates of indicated worms (and displayed in Supplementary Fig. 6a) were analysed by SDS-PAGE and immunoblotting against GFP and tubulin as loading control. Dots denote presence of indicated mutations, typical result from n = 3 biologically independent experiments. c Worms expressing the UFD substrate and the mutations indicated by dots were analysed as in (b), n = 3 biologically independent experiments. d same as (c) but worms are expressing the indicated mutations or a rescue construct for daf-16 either in the intestine (Pges-1::daf-16::GFP) or in neurons (Punc-119-daf-16::GFP), typical result of n = 3 biologically independent experiments. e same as (b) but indicated mutant worms were grown on control (ctrl) or tax-6 (RNAi) plates, n = 3 biologically independent experiments. f Schematic overview of experiments performed. UPS function becomes highly sensitive to the action of calcineurin signalling only when a neuronal signal mediated through gcy-8 (or absence of dyf-1) is present. g Indicated mutant worms expressing the UFD substrate were analysed at day one of adulthood by Western blotting against GFP and tubulin as loading control. h Quantification of worms displayed in (g), n = 3 biologically independent experiments. Column graphs: mean ± standard deviation, scatter plots: individual experiments, same colour: same experiment. 1-tailed unpaired Student’s t- test, *p ≤ 0.05, n.s. p > 0.05. dyf-1(mn335) versus dyf-1(mn335); ins-5(tm2560) p = 0.01114, dyf-1(mn335) daf-16(mu86) versus dyf-1(mn335) daf-16(mu86); ins-5(tm2560) p = 0.03790. Source data are provided as a Source data file.