STING negatively regulates canonical autophagy in Drosophila fat body. (A) RT-qPCR was performed to measure the knockdown efficiency of STING RNAi driven by Da-Gal4 or Arm-Gal4 at 28°C. Each data point was repeated three times with independent samples and normalized with ribosomal rp49 gene as an internal control. (B) Micrographs of WT or dSTING siRNA knockdown fat bodies from 92–94-h-old size-matched larva fed with or without CQ (3 mg/ml) for 24 h. Projections of three sections’ confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (C) Quantification of Atg8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats with or without CQ treatment. **, P ≤ 0.01; NS, not significant. (D) Micrographs of WT or STING−/− fat bodies from 92–94-h-old size-matched larva with or without CQ (3 mg/ml) for 24 h exposed to 20% sucrose solution to induce amino acid starvation. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (E) Quantification of Atg8a puncta in fat bodies averaged from five starved larvae from one representative experiment out of three repeats with or without CQ treatment. **, P ≤ 0.01; NS, not significant. (F) Micrographs of WT or dSTING KO fat bodies from 92–94-h-old size-matched larva fed with or without CQ (3 mg/ml) for 24 h. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (G) Quantification of Atg8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats with or without CQ treatment. ****, P ≤ 0.0001; NS, not significant. (H) Micrographs of fat bodies from 92–94-h-old size-matched larva fed. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. GFP-LAMP was detected by GFP fluorescence in fixed tissue. Scale bar, 20 µm. (I) Quantification of GFP-LAMP puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats. ***, P ≤ 0.001. (J) Quantification of ATG8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats. ***, P ≤ 0.001. (K) Micrographs of WT or dSTING knockdown fat bodies from 92–94-h-old size-matched larva fed or starved. Projections of three sections of confocal micrographs encompassing six to eight cells stained with Lysotracker. Scale bars are 20 µm. (L) Quantification of Lysotracker puncta in fat bodies averaged from five fed or starved larvae from one representative experiment out of three repeats. **, P ≤ 0.01. ST, STING.
Figure 1.

STING negatively regulates canonical autophagy in Drosophila fat body. (A) RT-qPCR was performed to measure the knockdown efficiency of STING RNAi driven by Da-Gal4 or Arm-Gal4 at 28°C. Each data point was repeated three times with independent samples and normalized with ribosomal rp49 gene as an internal control. (B) Micrographs of WT or dSTING siRNA knockdown fat bodies from 92–94-h-old size-matched larva fed with or without CQ (3 mg/ml) for 24 h. Projections of three sections’ confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (C) Quantification of Atg8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats with or without CQ treatment. **, P ≤ 0.01; NS, not significant. (D) Micrographs of WT or STING−/− fat bodies from 92–94-h-old size-matched larva with or without CQ (3 mg/ml) for 24 h exposed to 20% sucrose solution to induce amino acid starvation. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (E) Quantification of Atg8a puncta in fat bodies averaged from five starved larvae from one representative experiment out of three repeats with or without CQ treatment. **, P ≤ 0.01; NS, not significant. (F) Micrographs of WT or dSTING KO fat bodies from 92–94-h-old size-matched larva fed with or without CQ (3 mg/ml) for 24 h. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. Scale bar, 20 µm. (G) Quantification of Atg8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats with or without CQ treatment. ****, P ≤ 0.0001; NS, not significant. (H) Micrographs of fat bodies from 92–94-h-old size-matched larva fed. Projections of three sections of confocal micrographs encompassing six to eight cells stained for Atg8a. GFP-LAMP was detected by GFP fluorescence in fixed tissue. Scale bar, 20 µm. (I) Quantification of GFP-LAMP puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats. ***, P ≤ 0.001. (J) Quantification of ATG8a puncta in fat bodies averaged from five fed larvae from one representative experiment out of three repeats. ***, P ≤ 0.001. (K) Micrographs of WT or dSTING knockdown fat bodies from 92–94-h-old size-matched larva fed or starved. Projections of three sections of confocal micrographs encompassing six to eight cells stained with Lysotracker. Scale bars are 20 µm. (L) Quantification of Lysotracker puncta in fat bodies averaged from five fed or starved larvae from one representative experiment out of three repeats. **, P ≤ 0.01. ST, STING.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal