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. Author manuscript; available in PMC: 2023 Feb 11.
Published in final edited form as: Cell Rep. 2022 Dec 27;41(13):111875. doi: 10.1016/j.celrep.2022.111875

Figure 4. daf-2 inhibition promotes atfs-1-dependent mtDNA replication.

Figure 4.

(A) Fluorescence images of hsp-6pr::gfp and daf-2(e1370);hsp-6pr::gfp worms raised on control or cco-1(RNAi) at the L4 stage (scale bar, 0.1 mm).

(B) mtDNA quantification of wild-type, atfs-1(null), daf-2(e1370), and daf-2(e1370);atfs-1(null) worms, determined at the L4 stage (n = 5 ± SD, ****p < 0.0001, one way ANOVA with post hoc Tukey’s multiple comparisons test).

(C) mtDNA quantification of wild-type, atfs-1ΔNLS, daf-2(e1370), and daf-2(e1370);atfs-1DNLS worms determined at the L4 stage (n = 3 ± SD, ****p < 0.0001, one-way ANOVA with post hoc Tukey’s multiple comparison test).

(D) mtDNA quantification after ATFS-1 or control ChIP in wild-type and daf-2(e1370) worms at the L4 stage (n = 3 ± SD, **p = 0.0017, one-way ANOVA with post hoc Tukey’s multiple comparisons test).

(E) Images of wild-type, atfs-1(null), atfs-1ΔNLS, daf-2(e1370), daf-2(e1370);atfs-1(null), and daf-2(e1370);atfs-1ΔNLS worms under control and L1 arrest-fed conditions, obtained when control worms reached the L4 stage (scale bar, 0.5 mm).

(F) mtDNA quantification of wild-type, atfs-1(null), atfs-1ΔNLS, daf-2(e1370), daf-2(e1370);atfs-1(null), and daf-2(e1370);atfs-1ΔNLS worms under control conditions, as determined by qPCR at the L4 stage (n = 3 ± SD, *p = 0.132, one-way ANOVA with post hoc Tukey’s multiple comparisons test).

(G) mtDNA quantification of wild-type, atfs-1(null), atfs-1ΔNLS, daf-2(e1370), daf-2(e1370);atfs-1(null), and daf-2(e1370);atfs-1ΔNLS worms under L1 arrest conditions, as determined by qPCR when control worms reached the L4 stage (n = 3 ± SD, ****p < 0.0001, one-way ANOVA with post hoc Tukey’s multiple comparisons test).