Extended Data Fig. 2: Selective proteasomal regulation by MSTR-1 determines germ cell sex.
From: Noncanonical inheritance of phenotypic information by protein amyloids
(a) Schematic of the forward genetic screen for identifying suppressors of mstr germline feminization. ‘sdm’, suppressors of the double mstr worms, temporary nomenclature. Bar, 1 mm. (b) Brood sizes of the top suppressors of mstr feminization. Lines, mean (± S.D.). (c) Representative mapping plots of suppressors that were mapped to regions with mutations in proteasomal subunits but not validated by CRISPR/Cas9. Mapping data for all suppressors shown in b and c are provided in Source Data. Specific information on mapped mutations is listed in Supplementary Table 1. (d) Assessment of proteasomal function determined by expression of GFP tagged with uncleavable ubiquitin, Ub(G76V)::GFP. Plots, mean (± S.D.). ****P < 0.0001, ** P < 0.01, * P < 0.05. Two-way ANOVA with Bonferroni’s correction. Whole-worm difference in WT vs mstr at 20 °C is only significant when excluding pas-1 positive control from the analysis. Whole WT worms n = 3 20 °C, 9 P0 25 °C, 6 F1 25 °C worms. Whole mstr worms n = 9 20 °C, 10 P0 25 °C, 8 F1 25 °C worms. pas-1 RNAi treated worms: n = 3. WT germlines: n = 6 20 °C, 6 P0 25 °C, 4 F1 25 °C. mstr germlines: n = 8 20 °C, 7 P0 25 °C, 10 F1 25 °C. Exact P values are provided in Source Data. (e) Worms treated with proteasome inhibitor MG132 at L3 stage. Lines, mean (± S.D.). ****P < 0.0001, * P < 0.05, ns not significant. Percentages, percent of variation attributable to stated independent variable. Two-way ANOVA with Bonferroni’s correction. WT DMSO and MG132: n = 12 worms in each P0-F5 generation and treatment, 6 F6 worms in each treatment. mstr: n = 12 mstr worms in each generation and treatment. mstr; rpt-1 on DMSO: 5 P0, 6 F1, 6 F2, 8 F3, 7 F4, 8 F5, 8 F6 worms. mstr; rpt-1 on MG132: 6 P0-F3 each, 8 F4-6 each. Exact P values are provided in Source Data. (f) Expression of autophagy genes in mstr worms. Two-way ANOVA with Bonferroni’s correction. n = 3 WT and mstr worm populations. Numerical data and statistical values are available in source data.
