p62 aggregates sequester Atg7 in a LC3-dependent manner.

GFP-Atg7 (Aa), GFP-Atg7/Flag-LC3 (Ab), or GFP-Atg7^C572A/Flag-LC3 (Ac) vectors were co-transfected with mRFP-p62 into MEFwt cells for 48 h. GFP-Atg7 punctae were shown; mRFP-p62 and Flag-LC3 with GFP-Atg7 vectors (B) or with GFP-Atg7^C572A(C) were co-transfected into MEFwt cells for 48 h; mRFP-p62 and Flag-LC3A¹²⁰ with GFP-Atg7 vectors (D) or with GFP-Atg7^C572A (E) were co-transfected into MEFwt cells for 48 h. Flag-LC3, GFP-Atg7, or GFP-Atg7^C572A expression vectors were co-transfected into HEK293 cells as indicated for 48 h. Total cell lysates were used for IP by the Flag M2 antibody. Immunoblotting was conducted using the antibodies as indicated (F and G). Expressing vectors of GFP-Atg12 and mRFP-p62 were co-transfected into MEFwt cells for 48 h (H). All images were recorded at 48 h post-transfection. Scale bar = 25 micron. The inserts were enlarged to show the puncta in detail.