Uas yfp rab7

Flies were crossed, reared and kept on standard food (Meidi LLC) at 25°C. The following Drosophila strains used in this study had been previously generated by us (Z.H.’s lab): Hand-GFP (expressing green fluorescent protein in the nuclei of nephrocytes and cardiomyocytes; Han and Olson, 2005 (link); Huang et al., 2023 (link)), and Mhc-ANF-RFP (expressing ANF red fluorescent protein in muscle myosin heavy chain promoter-driven atrial natriuretic peptide-red fluorescent protein in muscle cells; Zhang et al., 2013 (link)). The following lines were obtained from the Bloomington Drosophila Stock Center (BDSC; Indiana University, IN): Dot-Gal4 (ID 6903; Ugt36A1 driver), tub-Gal4 (ID 30029; alphaTub84B driver), UAS-YFP-Rab5 (ID 24616), UAS-YFP-Rab7 (ID 23270), UAS-YFP-Rab11 (ID 9790), and UAS-mCherry-Atg8a (ID 37750). As control, w¹¹¹⁸ (BDSC; ID 3605) flies were used in the crosses.

Flies were reared on medium containing corn flour, sugar, yeast powder and agar along with antibacterial and antifungal agents. Flies were maintained at 25°C and 50% relative humidity. There was no internal illumination within the incubator, and the flies were subjected to light pulses of short duration only when the incubator door was opened. When required, flies were grown in an incubator with constant illumination from a white light source (intensity ~2000 lux).
The wild type used for all experiments was Red Oregon-R. GAL4-UAS system was used to drive expression of transgenic constructs. The following transgenic lines were obtained from the Bloomington Stock Center: UAS-GFP::Rab5 (B#43336),UAS-YFP::Rab7 (B#23270). UAS-garz^RNAi (V# 42140) was obtained from the Vienna Drosophila RNAi Center. Dicer;UAS-vps35^RNAi was obtained from Miklós Sass ( Eötvös Loránd University, Budapest, Hungary) and UAS-vps35::HA was obtained from Prof. Hugo Bellen (Baylor College of Medicine, Howard Hughes Medical Institute, Houston).