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. 2013 Jul 25;288(36):25760–25768. doi: 10.1074/jbc.M113.486258

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Analysis of the interaction of PhLP with Gβ and Gβγ subunits in cell culture. A, HEK 293 cells were co-transfected with the plasmids encoding the indicated epitope-tagged proteins and analyzed by pull-down with anti-HA or anti-FLAG-agarose after 48 h. B, representative blots showing captured Gγ₁-HA and its co-precipitated endogenous Gβ_{HEK 293}, Gβ₁-myc, PhLP-FLAG, and phosducin. C, representative blots showing captured PhLP-FLAG and its co-precipitated Gβ_{HEK 293}, Gβ₁-myc, Gγ₁-HA, and phosducin. D, the Gβ₁-myc and Gβ_{HEK 293} protein bands in C were quantified, and their values were normalized to that of PhLP-FLAG and expressed as a percentage fraction in lane 3 compared with lane 1; Gβ_{HEK 293} and Gβ₁-myc were reduced by 22 ± 10% (p = 0.094) and 29 ± 9% (p = 0.023). Error bars, S.E. with n = 5. Paired t test was applied.