(A) Schematic view of Atg9Gal4KO and Atg9d51mutations relative to the Atg9 transcripts. For the Atg9Gal4KO mutation, the complete Atg9 open reading frame was replaced with a Gal4 knock-in cassette. …
Quantification of lysotracker dots.
(A) LysoTracker Green staining reveals that developmental autophagy is strongly reduced in Atg9 mutant larval fat bodies, compared with controls. The number of lysotracker-positive dots in control …
Quantification of lysotracker dots and gastric caeca size.
(A) Both Atg7 and Atg9 mutant flies showed shortened lifespan compared with control (Log-rank test, p<0.001). Comparing with Atg7 mutant, Atg9 mutant flies exhibited dramatically shortened lifespan. …
Survival rate and climbing activity of control and Atg9 mutants.
(A) Atg9 mutant midguts are shortened and display enlargement in the posterior region (arrow). Scale bar: 500 μm. Quantification of adult midgut length and posterior midgut width of control and Atg9 …
Quantification of midgut length and width, cell size, total cell number, and PH3+, Delta+, Pros+ cells per gut.
(A) Representative images of Smurf flies. Smurf-, the blue dye is restricted to the proboscis and digestive tract. Smurf+, the dye is seen throughout the body due to intestinal barrier dysfunction. …
Percentage of Smurfs.
(A) Expression Atg9RNAi in ISCs, EBs and ECs with Dl-Gal4, Su(H)GBE-Gal4, or NP1-Gal4, respectively. Ablation of Atg9 in ECs, but not ISCs or EBs, caused enlarged cell size. n ≥ 25, ***p<0.001. (B) …
Quantification of cell size.
Temporal control of Atg9RNAi expression using the muscle-specific how-Gal4; Gal80ts inducible system. The flies were either maintained at 18°C throughout development or shifted to 29°C after …
Clonal depletion of Atg9RNAi (GFP positive cells) in larval wing and eye discs showed that Atg9 depletion did not affect cell size, compared with controls (GFP negative cells). Scale bar: 20 μm. …
(A) Systematic knock-down of Drosophila Atg1, Atg7, Atg9, Atg12, Atg13, Atg16, Atg17, Atg18 and Vps34 in adult midgut with the EC-specific driver NP1-Gal4; Gal80ts. The flies were either kept at …
Quantification of cell size.
(A) Quantification of Smurf flies showing that temporal knockdown of Atg1, Atg9, Atg13 and Atg17 resulted in intestinal barrier dysfunction in 30-day-old flies, compared with controls. n ≥ 40, data …
Survival rate and percentage of Smurfs.
(A) Temporal control of Atg genes RNAi expression using the Tub-Gal80ts; Tub-Ga4 inducible system. The flies were either maintained at 18°C throughout development or shifted to 29°C 1 day after egg …
Quantification of Atg8 dots.
(A) The adult midguts of denoted genotypes were dissected, lysed, and subjected to Western blot analysis with anti-Atg9, anti-Ref(2)P, anti-p-S6K, anti-p-4EBP and anti-tubulin antibodies. (B) …
Quantification of cell size.
(A) Quantification of Smurf flies showing that rapamycin treatment significantly decreased the number of Smurf-positive flies in aged Atg9 mutants. n ≥ 120, data are mean ±s.e.m., **p<0.01. ns, not …
Survival rate and percentage of Smurfs.
The Atg9RNAi-induced midgut defects (A) could be suppressed by the coexpression of InRRNAi (B), PI3KRNAi (C), or AktRNAi (D), whereas coexpression of PI3K (E) or Akt (F) dramatically enhanced Atg9RNA…
Quantification of cell size.
(A) Lysates of cells expressing Myc-Patj were incubated with GST or GST–Atg9-C (amino acids 668–845) in GST pull-down assays. The pull-down products and input Myc-Patj were analyzed by Western blots …
Quantification of cell size.
(A) Overexpression Atg9 or Patj could not suppress the Atg1RNAi-induced midgut defects. (B) Overexpression of Patj could not suppress the Atg17RNAi-induced midgut defects. The cell size of posterior …
Survival rate and quantification of cell size.
(A) S2 cells transfected with pWA-Gal4, pUAS-Flag-TSC2 and pUAS-GFP-Atg9 were subjected to immunoprecipitation with anti-Flag or anti-GFP antibody. The immunoprecipitates and total cell lysates were …
Quantification of fluorescent intensity and Western blots.
(A) S2 cells treated with dsRNAs targeting luciferase (Luc) or Atg1 were transfected with pWA-Gal4, pUAS-GFP-Atg9, pUAS-Myc-Patj, and pUAS-Flag-TSC2 and subjected to immunoprecipitation with …
Quantification of fluorescent intensity.
Adult flies fed with paraquat caused a marked increase of Atg8 puncta formation in midgut cells. Loss of Atg7 or Atg9 resulted in a marked decrease of paraquat-induced Atg8 puncta formation.