1. Structural and functional analysis of the GABARAP interaction motif (GIM)
GFP‐mCherry‐p62/SQSTM1 WT, T339V, L341V and T339V/L341V were overexpressed in HEK293 cells and immunoprecipitated using anti‐RFP beads and subjected to SDS–PAGE and Western blotting. Blots were probed for the presence of endogenous GABARAP and LC3B proteins. 3xFlag‐FIP200 WT, E703V, I705V, E703V/I705V and F702A/I705A were overexpressed in HEK293 cells and immunoprecipitated using anti‐Flag beads and subjected to SDS–PAGE and Western blotting. Blots were probed for the presence of endogenous GABARAP and LC3B proteins. GFP‐FUNDC1 WT, E19V, L21V, E19V/L21V and Y18A/L21A were overexpressed in HEK293 cells and either treated with vehicle only (DMSO) or 20 μM CCCP for 2 h, lysed and GFP‐FUNDC1 immunoprecipitated using anti‐GFP beads (or anti‐RFP beads as control) and subjected to SDS–PAGE and Western blotting. Blots were probed for the presence of endogenous GABARAP and LC3B proteins. Source data are available online for this figure.
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EMBO Rep, Volume: 18, Issue: 8, Pages: , First published: 27 June 2017, DOI: ( /embr )