Figure 5: DAF-16 activity in the F1 progeny of heat-shocked mothers is required for their enhanced stress resilience.

(A) Top: Experimental protocol to test the dependency of stress resilience in F1 progeny. Note:F1 progeny and not the P0 maternal germline were subjected to daf-16 or hsf-1 RNAi. Bottom: Stress resilience of F1 progeny from control non-heat shocked [NHS(Ctrl)], and heat-shocked, P0 mothers. Heat-shock: 5-minutes and 60-minutes. Control RNAi: L4440. n=3-10 experiments, 19.3±4.3 to 28.4±4.8 F1 progeny/experiment/condition.

(B) Stress resilience of F1 progeny from NHS(Ctrl) and heat-shocked P0 mothers. Heat-shock: 5-minute and 60-minute. P0 mothers were subjected to daf-16 germline RNAi. n=5-8 experiments, 24.4±2.7 to 35.2±2.0 F1 progeny/experiment/condition.

(C) Schematic of daf-2 gene regions analyzed for HSF-1 binding (5’ Far; −9071 to −8954), and H3K9me2 occupancy: a (−172 to −70), b (Intron 1: +3179 to +3279), c (Intron 2: +8046 to +8177) and d (Exon 9: +15719 to +15834).

(D) HSF-1 occupancy at the 5’-far region of daf-2 gene in NHS(Ctrl)] and heat-shocked wild-type and hsf-1 (sy441) P0 mothers. Heat-shock: 5-minute and 60-minute. n=3 experiments. Data normalized to NHS(Ctrl) wild-type and hsf-1(sy441) values respectively (also see Figure S7D).

(E) H3K9me2 occupancy at regions a, b, c, and d, depicted in C, in wild-type and hsf-1 (sy441) P0 mothers. Conditions: NHS(Ctrl), 2 hours after a 5-minute heat-shock (5min HS+rec), and immediately after a 60-minute heat-shock. n=3-4 experiments. H3K9me2 occupancy shown relative to NHS(Ctrl) wild-type and NHS(Ctrl) hsf-1 (sy441) values respectively. (also see Figure S7F).

A, B, D, E: Data show Mean ± Standard Error of the Mean. *: p<0.05, **: p<0.01, ***: p<0.001, ns: non-significant; ANOVA with Tukey’s correction, and Unpaired Student’s t-test.

See also Figure S7