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. 2010 Dec 8;286(7):5126–5135. doi: 10.1074/jbc.M110.180711

FIGURE 4.

FIGURE 4.

Mutually exclusive binding of Atgl and Abdh5 to Plin5. A, binding of Abhd5-Cherry suppresses binding of ECFP-Atgl to Plin5-EYFP. COS7 cells expressing Plin5-EYFP were fixed and permeabilized and then bound with ECFP-Atgl alone (top panel) or with Abhd5-Cherry (bottom panel). Images are from a representative experiment, and all capture parameters were equal for each fluorescence channel. Coincubation with Abhd5-Cherry significantly reduced binding of ECFP-Atgl as assessed by the regression analysis (p < 0.001). B, binding of ECFP-Atgl to Plin5-EYFP prevents binding of Abhd5-Cherry. COS7 cells coexpressing variable amounts of ECFP-Atgl and Plin5-EYFP were fixed and permeabilized and then bound with Abhd5-Cherry. A, B, and C are cells with variable amounts of bound ECFP-Atgl. Binding was assessed by the intensity of bound Abhd5-Cherry fluorescence as a function of Plin5 fluorescence (Plin5 Fluor.) and Atgl fluorescence (Atgl Fluor.) intensities. Binding was directly proportional to Plin 5 fluorescence (slope = +0.78) and inversely proportional to Atgl fluorescence (slope = −0.74). Bar, 10 μm.