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. Author manuscript; available in PMC: 2020 Jun 4.
Published in final edited form as: Nature. 2019 Dec 4;576(7786):301–305. doi: 10.1038/s41586-019-1814-y

Extended Data Figure 6. Effects of H3K4me3 down-regulation on heat shock response and endogenous redox state.

Extended Data Figure 6.

(a) ASH-2 and SET-2 transcript levels of N2jrIs2[Prpl-17::Grx1-roGFP2] worms treated with ash-2 or set-2 RNAi for 2 generations. n = 6 (ash-2) and n = 2 (set-2) independent experiments; unpaired t-test, two-sided. E.V., empty vector. (b) ASH-2 protein levels in N2jrIs2[Prpl-17::Grx1-roGFP2] worms treated with control RNAi or ash-2 RNAi for 2 generations using westernblot analysis. (c) H3K4me3 levels in N2jrIs2[Prpl-17::Grx1-roGFP2] worms treated with control RNAi, ash-2 RNAi or set-2 RNAi for 2 generations. (d) Transcript levels of selected heat shock genes after heat shock treatment of N2jrIs2[Prpl-17::Grx1-roGFP2] worms treated with the indicated RNAi. n = 3 independent experiments; one-way ANOVA with Bonferroni correction. (e) Transcript levels of selected heat shock genes in set-2 or wdr-5.1 mutants before and after heat shock treatment. n = 3 independent experiments; one-way ANOVA with Bonferroni correction. (f) ASH2L levels following ash2L siRNA treatment of HeLa cells. n = 2 independent experiments. (g) Grx1-roGFP2 ratios of L2 larval worms treated with ash-2 RNAi, set-2 RNAi, wdr-5.1 RNAi or the empty vector were measured using the BioSorter. n = 4 (ash-2, set-2) and n = 3 (wdr-5.1) independent sorting experiments; unpaired t-test, two-sided. (h) Representative survival curves of N2jrIs2[Prpl-17::Grx1-roGFP2] worms treated with ash-2 or set-2 RNAi for 2 generations, and treated with 1 mM PQ for 10 hours at the L2 larval stage. For n numbers, repetitions and statistics (log-rank), see Extended Data Table 4. Data in (a), (d), (e-g) represent mean ± SEM. For blot source images, see Supplementary Fig. 1 and 3.