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. 2022 Jan 28;34(4):1354–1374. doi: 10.1093/plcell/koac014

Figure 2.

Figure 2

ATL31 interacts with SYP61. A, Co-immunoprecipitation of GFP-SYP61 (56 kDa) with ATL31-FLAG or ATL31C143S-FLAG (43 kDa, indicated by arrowhead). Proteins were expressed in N. benthamiana leaves and immunoprecipitated with anti-FLAG antibody beads, followed by immunoblotting with anti-SYP61 and anti-FLAG antibodies. Asterisks, nonspecific bands and IgG heavy chain. B, Split ubiquitin yeast-two hybrid assays of ATL31-Cub and ATL31C143S-Cub with Nub-SYP61 or empty vector. Yeast cultures were diluted 1- or 10-fold (indicated by 1.0 or 0.1, respectively) and grown on solid medium with or without histidine (His). Cub, C-terminal half of ubiquitin; Nub, N-terminal half of ubiquitin. C, Co-immunoprecipitation of endogenous SYP61 with ATL31-GFP in Arabidopsis. Proteins were extracted from Arabidopsis plants expressing GFP (27 kDa, indicated by arrow) or ATL31-GFP (71 kDa, indicated by open arrowhead) and subjected to immunoprecipitation with anti-GFP antibody beads, followed by immunoblotting with anti-SYP61 and anti-GFP antibodies.