Loss of known autophagy regulators enhances p62 puncta formation cell-autonomously.

<p>Knockdown of Atg1 (A) or Tsc2 (B) increases p62 aggregate formation, while silencing of RpS8 results in a slight reduction of p62 dot number (C). Panel D shows statistical evaluation of the effect of RNAi and overexpression lines on p62 accumulation. Overexpressed p62-GFP forms multiple aggregates in control and Atg1 RNAi cells (panels E and F, respectively). Exposure times are indicated in the top right corner for panels E and F. Statistical evaluation of p62-GFP aggregate size and number in various RNAi and overexpression lines reveals changes that are difficult to interpret (G), but p62-GFP levels inferred from exposure times during image acquisition are qualitatively similar to data obtained with anti-p62 immunostaining (compare H to D). * indicates a significant difference (p<0.05), ** indicates a very significant difference (p<0.01), based on two-tailed two-sample unequal Student’s t tests in panels D, G, H. Scalebar in panel A equals 30 µm for panels A–C and E–F. <i>Genotypes are: (A) hsFlp; UAS-Dcr2/+; Act>CD2>Gal4, UAS-GFPnls/Atg1[JF02273]; (B) hsFlp; UAS-Dcr2/TSC2[KK103417]; Act>CD2>Gal4, UAS-GFPnls/+; (C) hsFlp; UAS-Dcr2/RpS8[KK106835]; Act>CD2>Gal4, UAS-GFPnls/+; (E) hsFlp; UAS-p62-GFP/+; Act>CD2>Gal4, UAS-Dcr2/+ (F) hsFlp; UAS-p62-GFP/+; Act>CD2>Gal4, UAS-Dcr2/Atg1[JF02273].</i>