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. 2019 Aug 13;17(8):e3000415. doi: 10.1371/journal.pbio.3000415

Fig 6. MDT-15 is required for maintaining HSF-1–dependent proteostasis at low temperatures.

Fig 6

(A) Fluorescence images of polyQ::YFP (Q35::YFP) transgenic worms in wild-type and mdt-15(tm2182) [mdt-15(-)] backgrounds at 15 °C. The images were captured at day 4 adult stage. Scale bar: 200 μm. (B) Quantification of the data shown in panel A (n ≥ 28 from three independent experiments, two-tailed Student t tests, **p < 0.01). These data are also used in Fig 7D for comparison. (C) Age-dependent changes in paralyzed polyQ::YFP transgenic worms in wild-type and mdt-15(-) backgrounds at 25 °C and 15 °C. mdt-15(-) mutations substantially accelerated the age-dependent paralysis at 15 °C. The transgenic worms in mdt-15(-) and wild-type backgrounds were shifted from 20 °C to indicated temperatures at L1 stage. p-values were calculated with log-rank test and were smaller than 0.0001 for wild-type versus mdt-15(-) animals both at 25 °C and at 15 °C. (D) Fluorescence images of hsp-16.1p::hsp-16.1::GFP [hsp-16.1::GFP] animals treated with mdt-15 RNAi or mdt-15/hsf-1 double RNAi at 15 °C. Scale bar: 200 μm. (E) Images of polyQ::YFP (Q40::YFP) transgenic worms treated with mdt-15 RNAi, hsf-1 RNAi, or mdt-15/hsf-1 double RNAi at 15 °C. The images were obtained by using L4 stage animals. Scale bar: 200 μm. (F) Quantification of the data shown in panel E (n ≥ 28 from three independent experiments, two-tailed Student’s t tests, **p < 0.01, ***p < 0.001). See S5 Table for statistical analysis and additional repeats of the paralysis assays and S7 Table for the standard deviations of box plot data. GFP, green fluorescent protein; HSF-1, heat shock factor 1; hsp, heat shock protein; MDT-15, Mediator 15; n.s., not significant; polyQ, polyglutamine; RNAi, RNA interference; YFP, yellow fluorescent protein.