Imaging for AgNO3 uptake was carried out with a Zeiss ApoTome.2 microscope using a 20× Plan-Apochromat 0.8 NA air objective. Confocal imaging for Dextran uptake, ANF-RFP, LysoTracker assays and anti-PDI was carried out using a Zeiss LSM900 microscope with a 20× Plan-Apochromat 0.8 NA air objective. Immunocytochemistry for anti-Pyd and anti-Ref(2)P were all carried out using confocal technology; a Zeiss LSM900 microscope with a 63× Plan-Apochromat 1.4 NA oil objective under Airyscan SR mode. Live-cell fluorescence imaging for the UAS-YFP-Rab5, UAS-YFP-Rab7, UAS-YFP-Rab11, and UAS-mCherry-Atg8a fly strains were all carried out using confocal technology; a Zeiss LSM900 microscope with a 63X Plan-Apochromat 1.4 NA oil objective under Airyscan SR mode. For quantitative comparison of intensities, common settings were chosen to avoid oversaturation. The results are presented as the mean±s.d. Statistical significance: *P<0.05, **P<0.01, ***P<0.001. ImageJ Software Version 1.52a was used for image processing.
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