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. Author manuscript; available in PMC: 2015 Feb 19.
Published in final edited form as: Nat Commun. 2014 Aug 19;5:4687. doi: 10.1038/ncomms5687

Figure 3. T35 is a BIN2 target site that controls CES protein stability.

Figure 3

(a) In vitro kinase assays using recombinant BIN2-GST and different GST-tagged CES variants. qM (quadruple mutant). CBB staining is shown as a loading control. (b) Representative fluorescence microscopic images of stomata of 14-day-old plants of 35S:CEST35A-YFP (line 2, upper two rows) or 35S:CEST35E-YFP (line 19, lower two rows) either untreated (upper rows) or treated for 24 h with 1 μM Brz followed by 2 h of 1 μM BL (lower rows). Scale bars: filter = 5 μm, magnified = 2 μm. (c) Immunoblotting of protein extracts of two-week old seedlings of the indicated lines using α-GFP antibody for CES-YFP detection. Upper panel: autoradiogram, lower panel: coomassie brilliant blue staining (CBB) as a loading control. (d) Immunoblotting of protein extracts of 7-day-old seedlings of 35S:CEST35E-YFP/16 and 35S:CESwt-YFP/32 in the presence or absence of MG132 (3 h treatment) using α-GFP antibody for CES-YFP detection. Bands were quantified with the image-quant software and the values are given. CBB staining is shown as a loading control.