Figure S4.

p62 over expression does not rescue aggresome clearance in Nrf1^KO cells. (A) Confirmation Western blot for p62 protein expression in HT22 Nrf1^KO and HT22 Nrf1^KO +p62^OE overexpression cells. α/β-Tubulin was used for loading control. (B) Representative images for autophagic flux in HT22 mCherry-GFP-LC3B Nrf1^KO and mCherry-GFP-LC3B Nrf1^KO +p62^OE cells treated for 20 h with DMSO or 200 nM BTZ. Images taken at 63×. Scale bar represents 15 μm. (C) Quantification of red and green puncta from BTZ treatment in panel B, signal is normalized to total cell number for each condition (Nrf1^KO n = 52 fields, Nrf1^KO + p62^OE n = 45 fields). (D) HT22 Nrf1^KO and Nrf1^KO+ p62^OE cells were treated with 50 nM CFZ for 20 h, then washed and either treated with 60 μM CQ or fresh complete media for a 20 h recovery (R). Western blot analysis of Ubiquitin after treatment and recovery, α/β-Tubulin was used for loading control. (n = 3 biological replicates). Error bars denote mean ± SD. Source data are available for this figure: SourceData FS4.