Extended Data Fig. 5: Loss of hpo-27 affects lysosomal degradation activity.
From: The HEAT repeat protein HPO-27 is a lysosome fission factor
a, Relative activity of cathepsin B and L in lysosomes purified from WT and hpo-27(tm5336) mutants. The amount of lysosomes in each strain was determined by western blot with an anti-ASP-4 antibody. b, Western blot analysis of NUC-1::CHERRY cleavage in WT and hpo-27(tm5336) at day 1. Quantification is shown in c. Four (a) and five (c) independent experiments. d-r, Confocal fluorescence images of WT, epg-5(tm3425), and hpo-27(tm5336) animals expressing GFP::LGG-1. Comma and 4-fold are embryonic stages; day 1 and day 3 are adult stages. LGG-1 puncta are indicated by white arrowheads. Quantifications are shown in s (n = 14 worms per strain). t-w, DIC images of the embryo (t, v) and germline (u, w) in WT and hpo-27(tm5336) mutants. Cell corpses are indicated by arrowheads. Quantifications are shown in x (n = 10 worms per strain), y (n = 15 worms per strain). Comma, 1.5 F, 2 F, 2.5 F, 3 F and 4 F (4-fold) are different embryonic stages. L4 is larval stage 4. In (a, c, s, x, y), data are shown as mean ± SD. Two-way ANOVA with Bonferroni post-test (s, x) or Student’s two-tailed unpaired t-test (a, b, y) was performed to compare mutant datasets with WT. P values are indicated. Scale bars: 5 µm. For gel source data, see Supplementary Fig. 1.
