Extended Data Fig. 5: ANC-1 preserves germline anatomy.
From: Nucleophagy delays aging and preserves germline immortality
a, Confocal microscopy imaging of day 2 WT and anc-1(RNAi)-mediated silenced FIB-1::GFP-expressing worm gonads grown at 25 °C. Scale bar, 10μm. b, Quantification of germ cell FIB-1::GFP nucleolar size and pixel intensity in WT and anc-1(RNAi)-mediated silenced worms from a. Mean ± s.d. ****P<0.0001 using one-way ANOVA; n ≥ 12 nucleoli. c, Epifluorescence microscopy imaging of GFP::ANC-1-expressing worm gonads. Arrows indicate distinct nucleoli visible by DIC microscopy, which are encircled by GFP::ANC-1. Scale bar, 10μm. d, DIC microscopy imaging of day 2 gonads of WT worms and gonads of worms subjected to RNAi-mediated silencing of the indicated genes grown at 25 °C after one generation and five generations. Scale bar, 20μm. e, Percentage of worm gonads grown at 25 °C whose proximal oocyte contains a nucleolus. Mean ± s.d. of three biological replicates. ****P<0.0001 using one-way ANOVA. f, Monitoring of protein synthesis rates by fluorescence recovery after photobleaching (FRAP) analysis. FRAP was performed on day 1 WT control, anc-1(RNAi)-, ife-2(RNAi)- or cycloheximide-treated (protein synthesis inhibitor) worms, expressing the reporter pife-2GFP. The ife-2 gene encodes the predominant eIF4E isoform in C. elegans that upon depletion blocks protein synthesis. The best-fit line illustrates the recovery rate, while the slope is the quantification of the recovery rate of a representative experiment.
