Fig. 1: Autophagy targets the nuclear membrane proteins nesprin-1 and nesprin-2.
From: Nucleophagy delays aging and preserves germline immortality
a, Confocal microscopy imaging of control (WT) and anc-1(RNAi)-mediated silenced worms of the indicated age (day 2 and day 13), expressing the plmn-1LMN-1::GFP reporter. The arrows indicate irregular nuclei (micronuclei, irregular shape, LMN-1 aggregation). Scale bar, 20 μm. b, Quantification of irregular nuclei per worm head and LMN-1::GFP fluorescence intensity in C. elegans head and body from a. The box plot represents the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. NS ≥ 0.05, ****P < 0.0001 using a two-way ANOVA; n ≥ 18 worm heads. c, Confocal microscopy imaging of day 2 adult worms expressing plgg-1GFP::LGG-1 and mKate2::ANC-1B in WT worms and on RNAi-mediated silencing of anc-1, lgg-1, rab-7 and daf-2. The arrows indicate GFP and mKate colocalization. Scale bar, 10 μm. d, Colocalization analysis (Pearson colocalization coefficient) between GFP::LGG-1 and mKate2::ANC-1B from c. The box plot represents the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. *P < 0.05, ****P < 0.0001 using a one-way ANOVA; n ≥ 18 worm midbody areas. e, Confocal microscopy imaging of day 2 adult worms expressing GFP::ANC-1B under anc-1, lgg-1 and rab-7(RNAi). Scale bar, 10 μm. f, Quantification of GFP::ANC-1 fluorescence intensity from e. The box plot represents the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. **P < 0.01, ****P < 0.0001 using a one-way ANOVA; n ≥ 27 worm midbody areas. g, Confocal microscopy imaging of day 2 adult worms expressing mKate2::ANC-1B after chloroquine, starvation, ultraviolet or combined treatments. Scale bar, 10 μm. h, Quantification of mKate2::ANC-1 fluorescent area and intensity from g. The box plot represents the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. **P < 0.01, ***P < 0.001, ****P < 0.0001 using a one-way ANOVA; n = 25 worm midbody areas. i, Western blot analysis of the nesprin-2 isoform ε1, p62 and LC3 from E14.5 MEF lysates under control, BafA1, starvation and BafA1/starvation conditions.
