1. IIS‐dfoxo‐dependent regulation of proteasome activity in the gut enhances proteome maintenance
IIS‐dfoxo‐dependent regulation of proteasome activity in the gut enhances proteome maintenance Tissue‐specific average log2 fold change of IIS‐regulated proteins associated to the proteasome.Western blot and quantification analysis of Rpt6 in wild‐type (wDah) (n = 8), dfoxoΔ94 mutants (n = 3), mNSC‐ablated flies (InsP3‐Gal4/UAS‐rpr) (n = 7) and mNSC‐ablated flies lacking dfoxo (InsP3‐Gal4/UAS‐rpr, dfoxoΔ94) (n = 4), normalized to protein loading and significance determined by one‐way ANOVA.Assessment of proteasome assembly by in‐gel caspase‐like (LLE) proteasome activity in mNSC‐ablated fly gut and control flies (wDah). Quantification shows the activity ratio 26S/20S (n = 5), significance determined by t‐test.Tissue‐specific caspase‐like activity was assessed using fluorogenic substrate (LLE‐AMC, Enzo Life Science) in wild‐type (wDah; n = head 14, fat body 5, gut 8, thorax 8), dfoxoΔ94 mutants (n = head 9, fat body 5, gut 7, thorax 8), mNSC‐ablated flies (InsP3‐Gal4/UAS‐rpr; n = head 14, fat body 5, gut 7, thorax 9) and mNSC‐ablated flies lacking dfoxo (InsP3‐Gal4/UAS‐rpr, dfoxoΔ94; n = head 7, fat body 5, gut 9, thorax 9), significance determined by two‐way ANOVA and post hoc pairwise tests.Proteasomal caspase‐like activity of an independent IIS mutant (dilp2‐3,5, grey) compared to controls (wDah) (gut n = 9, fat body n = 7). Statistical significance was determined by t‐test.Representative Western blot showing tissue‐specific levels of K48‐linked poly‐ubiquitin and their quantification (head, thorax and fat body Westerns performed in quadruplicate). For gut samples wild‐type (wDah) and dfoxoΔ94 mutants n = 11, InsP3‐Gal4/UAS‐rpr n = 12 and InsP3‐Gal4/UAS‐rpr, dfoxoΔ94n = 10, significance determined by two‐way ANOVA and post hoc pairwise tests. Data information: *P < 0.05, **P < 0.01, ***P <  All error bars show SEM.
Luke S Tain et al. Mol Syst Biol 2017;13:939
© as stated in the article, figure or figure legend