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. 2020 Mar 13;117(13):7482–7493. doi: 10.1073/pnas.1918539117

Fig. 4.

Fig. 4.

Direct interaction between COST1 and ATG8e. (A) Colocalization of COST1-YFP with mCherry-ATG8e and mCherry-NBR1. (Scale bar, 10 µm.) (B) Split luciferase analysis of the interaction between COST1-nLUC and cLUC-ATG8e. Different combinations of GFP-nLUC and cLUC-GFP with and without COST1 or ATG8e were used as negative controls. (C) GST pull-down assay between GST-COST1 and His-ATG8e. GST alone was used as negative control. (D) Co-IP of COST1-Flag with GFP-ATG8e. Agrobacterium-mediated coinfiltrations were carried out in tobacco leaves with combinations of 35S:GFP and 35S:COST1-Flag, and 35S:GFP-ATG8e and 35S:COST1-Flag. After 2 d of incubation, leaves were ground in liquid nitrogen and proteins immunoprecipitated with GFP-Trap. The immunoblot was probed with anti-GFP and anti-Flag antibodies. Leaves expressing 35S:GFP alone were used as a negative control.