Fig. 5: Pre-NRC starvation causes proteasome dysfunction.
From: Nutrient status alters developmental fates via a switch in mitochondrial homeodynamics
A Images of proteasomal activity (FK2 or CL1-GFP) in PG cells after 0 h, 3 h, 6 h and 12 h starvation from 72 h AEL. FK2, red; CL1-GFP, green. Magnified images of the highlighted areas (yellow dotted box) are shown. FK2: from left to right, n = 16, 12, 20 and 36, respectively. CL1-GFP: From left to right, n = 20, 33, 18 and 23, respectively. B, C Proteasomal activity visualized by FK2 (B, red) or CL1-GFP (C, green) was assessed in PG cells of phm > w1118, phm > Torso-RNAi, phm > RasDN, phm > InR-RNAi, phm > Dp110DN, phm > Torso + InR-RNAi, phm > TORDN and phm > TSC1/2 under control of Gal80ts. FK2: from left to right, n = 11, 12, 28, 11, 12, 15, 8 and 8, respectively. CL1-GFP: From left to right, n = 15, 26, 30, 26, 27, 7, 28 and 23, respectively. Magnified images of the highlighted areas (yellow dotted box) are shown. D, E Proteasomal activity visualized by FK2 (red) was assessed in PG cells of D phm > w1118 (n = 7), phm > RasV12 (n = 18), phm > Dp110CA (n = 9), phm > Rheb (n = 9), E phm > w1118 (n = 3), phm > Rpt3-RNAi (n = 3), phm > Rpn11-RNAi (n = 3), and phm > prosβ1-RNAi (n = 3). Magnified images of the highlighted areas (yellow dotted box) are shown. F, G Representative images of mitophagy (F, 561 nm, red; 405 nm, green) and mitochondria (G, green) in PG cells (132 h AEL) with knockdown of Rpt3, Rpn11 and prosβ1. F n = 17 (Control), 18 (Rpt3-RNAi), 11 (Rpn11-RNAi), 6 (Prosβ1-RNAi). G n = 16 (Control), 19 (Rpt3-RNAi), 19 (Rpn11-RNAi), 12 (Prosβ1-RNAi). H, I Images of proteasomal activity (FK2 antibody, red) H or CL1-GFP (green) I in PG cells of controls and PG-expressing ParkinCA under 29 °C. Magnified images of the highlighted areas (yellow dotted box) are shown. H n = 14 (Control), 9 (ParkinCA). J Proteasomal activity visualized by FK2 (red) was assessed in PG cells of phm > w1118 (n = 17), phm > COX6B-RNAi (n = 12), phm > Neb-cGP-RNAi (n = 14), phm > TrpRS-m-RNAi (n = 23), phm > ATPsynC-RNAi (n = 15), phm > mRPL47-RNAi (n = 4), phm > Tom20-RNAi (n = 15), and phm > mge-RNAi (n = 15). Magnified images of the highlighted areas (yellow dotted box) are shown. The genotypes and statistical data are provided in Supplementary Data 1.
