Figure 5. scav-3 mutants accumulate damaged lysosomes. (A–G) Confocal fluorescent images of the hypodermis in wild-type (WT; A–A″) and scav-3(qx193) (B–B″ and C–G) adults expressing NUC-1::mCherry and/or sfGFP::Gal3. sfGFP::Gal3-positive structures contain faint NUC-1::mCherry fluorescence (B–B″, circles). A ring of sfGFP::Gal3 surrounding a bright NUC-1::mCherry punctum in scav-3(qx193) is indicated by the arrows. (H) The percentage of lysosomes that contain strong mCherry fluorescence and are negative for GFP::Gal3 (intact) and those that contain faint mCherry fluorescence and are positive for GFP::Gal3 (damaged) was quantified in wild type and scav-3(qx193) expressing sfGFP::Gal3 and NUC-1::mCherry. At least 15 animals were scored in each strain, and data are shown as mean ± SD. (I) Confocal fluorescent images and linescan analysis of lysosomes in scav-3(qx193) adults expressing sfGFP::Gal3 and NUC-1::mCherry. sfGFP::Gal3-positive structures contain faint NUC-1::mCherry fluorescence, whereas lysosomes with bright NUC-1::mCherry lacked sfGFP::Gal3. At least 10 lysosomes were examined, and the representative result is shown. (J–K″) Confocal fluorescent images of the hypodermis in wild-type (J–J″) and scav-3(qx193) (K–K″) adults expressing sfGFP::Gal3 and stained by LysoTracker red. Arrows and arrowheads indicate sfGFP::Gal3-positive and LysoTracker red–positive structures, respectively. (L) Confocal fluorescent images and linescan analysis of lysosomes in scav-3(qx193) adults expressing sfGFP::Gal3 and stained by LysoTracker red. sfGFP::Gal3-positive structures were not stained by LysoTracker red, whereas sfGFP::Gal3 was absent from LysoTracker red–positive vesicles. At least 10 lysosomes were examined, and the representative result is shown. (M) The available acid phosphatase activity in suspensions of wild-type and scav-3(qx193) lysosomes was determined, and the mean percentage of total activity (obtained in the presence of 0.1% Triton X-100) is shown. Data were compared with the unpaired t test. *, P < 0.05. Bars, 5 µm.
Figure 5.

scav-3 mutants accumulate damaged lysosomes. (A–G) Confocal fluorescent images of the hypodermis in wild-type (WT; A–A″) and scav-3(qx193) (B–B″ and C–G) adults expressing NUC-1::mCherry and/or sfGFP::Gal3. sfGFP::Gal3-positive structures contain faint NUC-1::mCherry fluorescence (B–B″, circles). A ring of sfGFP::Gal3 surrounding a bright NUC-1::mCherry punctum in scav-3(qx193) is indicated by the arrows. (H) The percentage of lysosomes that contain strong mCherry fluorescence and are negative for GFP::Gal3 (intact) and those that contain faint mCherry fluorescence and are positive for GFP::Gal3 (damaged) was quantified in wild type and scav-3(qx193) expressing sfGFP::Gal3 and NUC-1::mCherry. At least 15 animals were scored in each strain, and data are shown as mean ± SD. (I) Confocal fluorescent images and linescan analysis of lysosomes in scav-3(qx193) adults expressing sfGFP::Gal3 and NUC-1::mCherry. sfGFP::Gal3-positive structures contain faint NUC-1::mCherry fluorescence, whereas lysosomes with bright NUC-1::mCherry lacked sfGFP::Gal3. At least 10 lysosomes were examined, and the representative result is shown. (J–K″) Confocal fluorescent images of the hypodermis in wild-type (J–J″) and scav-3(qx193) (K–K″) adults expressing sfGFP::Gal3 and stained by LysoTracker red. Arrows and arrowheads indicate sfGFP::Gal3-positive and LysoTracker red–positive structures, respectively. (L) Confocal fluorescent images and linescan analysis of lysosomes in scav-3(qx193) adults expressing sfGFP::Gal3 and stained by LysoTracker red. sfGFP::Gal3-positive structures were not stained by LysoTracker red, whereas sfGFP::Gal3 was absent from LysoTracker red–positive vesicles. At least 10 lysosomes were examined, and the representative result is shown. (M) The available acid phosphatase activity in suspensions of wild-type and scav-3(qx193) lysosomes was determined, and the mean percentage of total activity (obtained in the presence of 0.1% Triton X-100) is shown. Data were compared with the unpaired t test. *, P < 0.05. Bars, 5 µm.

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