Extended Data Fig. 1: hpo-27 mutants contain extensive lysosomal tubules.
From: The HEAT repeat protein HPO-27 is a lysosome fission factor
a-r, Confocal fluorescence images of the hypodermis in day 2 adult worms carrying different hpo-27 mutant alleles (a-p), or treated with hpo-27 RNAi (q, r). The worms were expressing NUC-1::CHERRY and AJM-1::GFP to label lysosomes and lateral seam cells, respectively. hpo-27 mutants contain extensive lysosomal tubules and greatly diminished NUC-1::CHERRY fluorescence in hyp7 cells. The percentage of worms with the representative lysosomal pattern is shown at the lower left corner in each panel. s, qPCR analysis of hpo-27 gene expression in control RNAi and hpo-27 RNAi worms. Three independent experiments were performed, and data are shown as mean ± SD. Student’s two-tailed unpaired t-test was performed to compare hpo-27 RNAi with control RNAi. P values are indicated. t-v”, Confocal fluorescence images of the hypodermis in wild type (WT) (t-t”) and hpo-27(tm5336) mutant adults (u-v”) carrying NUC-1::CHERRY and Psemo-1GFP (n = 15 worms in each strain). Lysosomes labeled by NUC-1::CHERRY are seen in both hyp7 cells positive for GFP and lateral seam cells that lack GFP expression (arrows). In hpo-27 mutants, extensive lysosomal tubules are present in hyp7 and seam cells, and NUC-1::CHERRY fluorescence is diminished in hyp7 but not seam cells. w, Cloning of hpo-27. The hpo-27 gene structure is shown with filled boxes representing exons, thin lines indicating introns, and gray bars designating 3’ and 5’ UTRs. The arrow delineates the direction of transcription. x, Schematic diagram showing the organization of the HPO-27 protein. The mutation sites identified in all the hpo-27 alleles and the region deleted in the tm5336 allele are indicated. Gray boxes indicate the HEAT motifs. Scale bars: 5 µm.
