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. 2017 Nov 16;6:e29338. doi: 10.7554/eLife.29338

Figure 4. Loss of Atg9 leads to enlarged enterocytes.

(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) Temporal control of Atg9RNAi expression using the Gal80ts; Tub-Gal4 inducible system. The flies were either maintained at 18°C throughout development or shifted to 29°C after eclosion for 5 days to inactivate Gal80ts and enable expression of the RNAi targeting Atg9. (C) Clonal analysis in adult midgut using Flp-FRT-mediated recombination revealed that Atg9d51 mutant cells (marked by lack of GFP and Atg9 expression) are larger than the controls (GFP-positive cells). n ≥ 17, ***p<0.001. (D) MARCM analysis showed that the enlarged Atg9d51 mutant cells (marked by GFP) are Pdm1 positive EC cells. n ≥ 21, ***p<0.001. Scale bar: 20 μm. Genotypes: (C) hsFLP; FRT42D Ubi-GFP/FRT42D Atg9d51 (D) hsFLP; FRT42D tubGal80/FRT42D Atg9d51; Tub-Gal4/UAS-mCD8GFP.

Figure 4—source data 1. Quantification of cell size.
elife-29338-fig4-data1.xlsx (10.4KB, xlsx)
DOI: 10.7554/eLife.29338.015

Figure 4.

Figure 4—figure supplement 1. Atg9 depletion in visceral muscle does not cause any observable defects in the midgut.

Figure 4—figure supplement 1.

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 eclosion for 5 days to inactivate Gal80ts and enable expression of the RNAi targeting Atg9. Scale bar: 20 µm.
Figure 4—figure supplement 2. Atg9 depletion does not affect cell size of larval imaginal discs.

Figure 4—figure supplement 2.

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. Genotypes: hsflp; Act-CD2-Gal4-UAS-GFP-Atg8a/UAS-Atg9RNAi.