Extended Data Fig. 3: Loss of HPO-27 affects lysosome morphology in intestine and body wall muscle cells.
From: The HEAT repeat protein HPO-27 is a lysosome fission factor
a-d, Confocal fluorescence images of the intestine and body wall muscle cells in WT and hpo-27(tm5336) adult worms expressing LAAT-1::GFP. Arrowheads indicate tubular lysosomes. Quantifications are shown in e, f, h (n = 14 worms per strain) and g (n = 15 worms per strain). Data are shown as mean ± SD. Two-way ANOVA with Bonferroni post-test (e, f, h) or one-way ANOVA with Tukey’s multiple comparison test (g) was performed to compare mutant datasets with WT. P values are indicated. i-q, Confocal fluorescence images of the hypodermis in WT and hpo-27(tm5336) worms at adult day 2 expressing AJM-1::GFP (which stains the seam cell boundary) and RNST-2::CHERRY (i-k) or SCAV-3::tagRFPt (l-n), or carrying Pcol-12LMP-1::RFP which is expressed in hyp7 but not seam cells (o-q). The percentage of worms with the representative lysosomal pattern is shown at the lower left corner in each panel. In (q), the DIC image is merged to show a hyp7 cell with invisible LMP-1::RFP. r-v’, Confocal fluorescence time-lapse images of the hypodermis in adult hpo-27(tm5336) worms co-expressing RNST-2::CHERRY and SCAV-3::GFP. The lysosomal tubular networks are clearly observed at “0 min”, but they gradually collapse. White arrowheads indicate the lysosomal tubules that gradually disappear. The lysosomal membrane protein SCAV-3::GFP labels tubular structures, which collapse to form small vesicles (yellow arrowhead) and eventually became diffuse. Scale bars: 5 µm.
