The loss of LRPPRC function induces the mitochondrial unfolded protein response

Figure 3.Inactivation of mma-1 by RNAi in C. elegans induces ATFS-1-dependent UPR^{mt}. Western analysis of (A) P_{hsp-6}GFP (mitochondrial Hsp70), (B) P_{hsp-60}GFP (mitochondrial Hsp60) or (C) P_{hsp-4}GFP (ER BiP) reporter strains treated with mock(RNAi), mma-1(RNAi) 1:5 dil. (diluted 1:5 (v/v) with mock(RNAi)) or spg-7(RNAi) 1:5 dil. (diluted 1:5 (v/v) with mock(RNAi)). Ratios of P_{hsp-6}GFP/Tubulin, P_{hsp-60}GFP/Tubulin and P_{hsp-4}GFP/Tubulin relative to the mock(RNAi) treated animals are indicated (n = 5 for P_{hsp-6}GFP + mma-1(RNAi); n = 7 for P_{hsp-6}GFP + spg-7(RNAi); n = 6 for P_{hsp-60}GFP + mma-1(RNAi); n = 9 for P_{hsp-60}GFP + spg-1(RNAi); n = 5 for P_{hsp-4}GFP). Western analysis of the effect of mma-1(RNAi) on endogenous (D) HSP-6 or (E) HSP-60 protein level. Ratios of HSP-6/Tubulin and HSP-60/Tubulin relative to the mock(RNAi) treated animals are indicated (n = 8 for HSP-6 and n = 10 for HSP-60). (F) Western analysis of the effect of ATFS-1 on mma-1(RNAi)-induced UPR^{mt}. mma-1(RNAi) or spg-7(RNAi) were diluted either with mock(RNAi) or atfs-1(RNAi). Relative ratios of P_{hsp-6}GFP/Tubulin are indicated (n = 7 for mma-1(RNAi) and n = 10 for spg-7(RNAi)). (G) Western analysis of the effect of HAF-1 on mma-1(RNAi)-induced UPR^{mt}. Wild-type P_{hsp-6}GFP reporter strain (+/+) or P_{hsp-6}GFP reporter strain carrying the haf-1(ok705) loss-of-function mutation were analyzed. The relative ratios of P_{hsp-6}GFP/Tubulin are indicated (n = 5 for mma-1(RNAi) and n = 6 for spg-7(RNAi)) (For all panels, average values are shown and error bars indicate s.d.; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 by one sample t-test).