Fig 7.

Fau interacts with p62 in vivo. (A), Fau, p62, and ubiquitin conjugates are colocalizing in cells with or without undergoing autophagy. RAW264.7 cells were subjected to autophagic induction by starvation for 2 h and immunostained for endogeneous Fau, p62, and ubiquitin conjugates. (B), Fau is found in the same immune complexes with p62. RAW264.7 cells were infected with mycobacteria for 1 h, washed, and harvested. Cell lysates were prepared and subjected for immunoprecipitation experiments and the immune complexes were analyzed by Western blotting with indicated antibodies. 100 μg of input also analyzed in parallel. (C) The ubiquitin associate domain (UBA) of p62 interacts with Fau in vivo. P62 full-length or PB1 domain containing R21A/D69A mutation (to prevent oligomerization) or UBA domain tagged with GFP plasmids were tranfected into RAW264.7 cells. 48 h after transfection, cells were subjected for immunoprecipitation and the immune complexes and cell lysates were analyzed by immunoblotting.