Figure 5.

Autophagy inhibition in Ras-activated tissues results in cell- and non-cell-autonomous effects on tissue growth. (a) Three-dimensional (3D) reconstruction of Ras^V12Atg^RNAi and control eye-antennal mosaic discs from L3 wandering larvae. Mutant clones are GFP+ and tissues were counterstained with DAPI (blue). (b) In discs expressing both Ras^V12 and Atg1^RNAi in clones, the proportion of GFP tissue was significantly higher than in the control Ras^V12 mRFP samples (49.4±2.4%, compared to 38.9±1.2%, P=0.00028). This trend was also observed in Ras^V12 Atg8a^RNAi, although the difference was not significantly different (42.9±2.3%, P=0.068). (c) Non-cell-autonomous proliferation in wild-type tissue (non-GFP) surrounding Ras^V12GFP, Ras^V12 Atg1^RNAi or Ras^V12Atg8a^RNAi clones (GFP+), as seen by EdU incorporation. (d) Non-cell-autonomous caspase activation in wild-type tissue (non-GFP) surrounding Ras^V12Atg1^RNAi or Ras^V12Atg8a^RNAi clones (GFP+), as seen by Dcp1 staining. Arrows indicate the apoptotic wave before the MF. (e) Apoptosis is confirmed by TUNEL assay around clones expressing Ras^V12 and Atg8a-RNAi. (f') Quantification of data in (d). (g) Quantification of data in (e). Error bars=s.e.m. Statistics: one-way ANOVA with Tukey's multiple correction test. Scale bars: 50 μm.