Fig. 8.

Inhibition of polyubiquitination decreases uptake of the proteasome by autophagosomes. (A) (i) GFP-LC3B–expressing HeLa cells were transiently transfected with p62. At 24 h later, the cells were infected with adenoviruses coding for either WT HA-Ub or HA-K0-Ub. Cells were split and were either left untreated or starved for amino acids for 1 h. The proteasome was immunoprecipitated from cell lysates, and the precipitates were resolved and blotted with anti-p62, anti-LC3B, anti-α6, and anti-HA. (ii) Expression of WT Ub and K0-Ub (Upper) and p62 (Middle) was detected in whole-cell lysates (WCL). (B) Immunofluorescent staining with anti-α6 of GFP-LC3B–expressing HeLa cells infected with WT Ub (Upper) or K0-Ub (Lower) was performed after 4 h of amino acid starvation. (Scale bars: 20 μm.)