Figure 2.

Loss of Hsp83 function leads to an increase in autophagy and cell death features in vivo. (A) Mid-stage egg chambers (MSECS) scored as being TUNEL positive, LysoTracker® Red (LTR)-positive or as having condensed degenerating nurse cell nuclei by DAPI (MSDEC) with percentages reported according to their genotype. At least 50 MSECs were counted per genotype (n = 3). Error bars represent ± SEM and statistical significance was determined using one-way ANOVA with a Bonferroni post-test and compared with w¹¹¹⁸, ***P < 0.001, ****P < 0.0001. (B, C) Representative MSDECs and nondegenerating MSECs stained with LTR, TUNEL and DAPI, scale bars: 25 μm. (B) MSDEC from an Hsp83^e6A/6–55 ovariole that scored positive for LTR and TUNEL staining. (C) Nondegenerating MSEC from Hsp83^6–55/+ scored as negative for LTR and TUNEL (D to I) MSECs were scored from flies expressing GFP-mCherry-Atg8a in the germline (UAsp-GFP-mCherry-DrAtg8a with single copies of the drivers otu-GAL4 and NGT-GAL4). Hsp83^e6D/+ and Hsp83^6–55/+, Hsp83^e6A/+ and Hsp83^6–55/+ from the same cross were analyzed together. (D) Percentage of MSDECs for the indicated genotypes is represented on the graph and reflects the mean of at least 100 MSECs scored per genotype (n = 3). Error bars represent ± SEM and statistical significance was determined using a 2-tailed Student t test, *P < 0.05. (E) Flies expressing GFP-mCherry-Atg8a in the germline were scored as either having more than 5 autolysosomes or less than or equal to 5 autolysosomes. The percentages shown reflect the mean of at least 100 MSECs scored per genotype (n = 3). Error bars represent ± SEM and statistical significance was determined using a 2-tailed Student t test, **P < 0.01. (F) to I) Representative images of MSECs expressing the construct GFP-mCherry-Atg8a, scale bar = 25μm. MSDECS found in (F) Hsp83^{e6D/6–55 flies} and (G) Hsp83^{e6D or 6-55/+}. (H, I) Examples of nondegenerating MSECs from (H) Hsp83^{e6D/6–55 flies} and (I) Hsp83^{e6D or 6-55/+}.