Figure 2.

HEC-23 impairs lysosomal degradation and increases the number of cell corpses in the C. elegans germline. (A) Representative images of TR-BSA localization in LMP-1::GFP-positive lysosomes following HEC-23 treatment. (B–D) Time-course analysis of ssGFP signals in DMSO- and HEC-23-treated coelomocytes. Expression of ssGFP under the control of a heat-shock promoter was induced at 33°C for 30 min, and the uptake and degradation of ssGFP in coelomocytes were monitored at the indicated time points (B). The dashed circles indicate HEC-23-enlarged lysosomes. Quantifications are shown in (C and D). (E–H) Images (E and F) and quantification (G and H) of HEC-23-induced germ cell corpses labeled with CED-1::GFP (E) or GFP::moesin (F) in the C. elegans germline. Arrows indicate cell corpses. Scale bars, 20 μm. (I) Quantification of HEC-23-induced germ cell corpses in animals at the indicated adult ages. 30 animals were scored for each time point. (J and K) Quantification of HEC-23-induced germ cell corpses in ced-3 (I) and ced-4 (J) loss-of-function mutants. (L) Durations of germ cell corpses in DMSO- and HEC-23 (100 μmol/L)-treated worms. Cell corpses from >30 worms were analyzed. For all quantifications, data (mean ± SEM) were from 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant