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. 2013 Sep 4;8(9):e73229. doi: 10.1371/journal.pone.0073229

Figure 3. p62 aggregates do not sequester Atg4B.

Figure 3

GFP-Atg4B and mRFP-p62 expression plasmids (A) or with Flag-LC3 (B) were co-transfected into MEFwt cells for 24h. GFP-Atg4BC74A and mRFP-p62 with Flag-LC3 vectors (C) or with Flag-LC3A120 vector (D) were co-transfected into MEFwt cells for 24 h. GFP-Atg4B or GFP-Atg4BC74A expression vector was co-transfected with Flag-LC3 (E) or Flag-LC3A120 (F) into HEK293 cells for 48 h. Immunoprecipitation (IP) was performed using total cell lysates. Immunoblotting was conducted using the antibodies as indicated. (G) mRFP-p62, GFP-Atg4BC74A and Flag-LC3 expression vectors were co-transfected into MEFwt cells for 24 h. Colocalization of Flag-LC3 within mRFP-p62 aggregates were detected by immunofluorescence staining (IF) using Flag M2 antibody. Fluorescence images presented in this study were representative of at least three independent experiments. Scale bar = 25micron.