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. Author manuscript; available in PMC: 2024 Sep 5.
Published in final edited form as: Autophagy Rep. 2023 Sep 5;2(1):2252307. doi: 10.1080/27694127.2023.2252307

Figure 5: Downregulation of M1BP triggers cell death.

Figure 5:

Eye-antennal imaginal discs stained for pan-neuronal marker Elav (red), Dlg (blue), a membrane specific marker, to mark outline of the tissue and (A-D) hid5’F-WT-GFP (green). Eye antennal imaginal disc showing split channel for (A’-D’) hid5’F-WT-GFP for (A, A’, B, B’) ey-Gal4 and (C, C’, D, D’) ey>M1BPRNAi in (A, A’, C, C’) 20X and (B, B’, D, D’) 40X. (E) Quantification of hid5’F-WT-GFP intensity using Fiji/ImageJ software (NIH) using standard 100X100 pixel ROI. ROI is shown as yellow dashed boxes. Caspase-dependent cell death was blocked in the entire eye (F, K, K’, O, O’) ey>hidRNAi, (H, M, M’, Q, Q’) ey>hidΔN14. Blocking hid mediated apoptotic cell death in ey>M1BPRNAi background (G, L, L’, P, P’) ey>M1BPRNAi+hidRNAi, (I, N, N’, R, R’) ey>M1BPRNAi+hidΔN14 shows rescue when compared to ey>M1BPRNAi. (J) Graphical representation of adult eye phenotype frequency. The genotypes depicted in the graph are 1: ey-Gal4, 2: ey>M1BPRNAi, 3: ey>M1BPRNAi+hidRNAi and 4: ey>M1BPRNAi+hidΔN14. Statistical significance with eyGal4 and ey>M1BPRNAi are depicted using # and * respectively. Eye antennal imaginal discs of indicated genotypes stained for (K-N) Wg (green), (O-R) pJNK (green) and pan-neuronal marker Elav (red). Eye-antennal imaginal discs showing split channel for (K’-N’) Wg and (O’-R’) pJNK staining. (S) pJNK intensity quantification using Fiji/ImageJ software (NIH). The genotypes depicted in the graph are 1: ey-Gal4, 2: ey>M1BPRNAi, 3: ey>M1BPRNAi+hidRNAi and 4: ey>M1BPRNAi+hidΔN14. Quantification was performed using standard 100X100pixels ROI. ROI is shown as yellow dashed boxes. Graphs were plotted with mean +/− SEM. Statistical significance in each graph is shown by p-value: ****p<0.0001, ***p<0.001; **p<0.01; *p<0.05. The orientation of all imaginal discs is identical with posterior to the left and dorsal up. The magnification of all eye-antennal imaginal discs is 20X and adult eyes are 10X unless specified.