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Figure 4 | Journal of Molecular Signaling

Figure 4

From: Mutations on the Switch III region and the alpha3 helix of Galpha16 differentially affect receptor coupling and regulation of downstream effectors

Figure 4

The alanine mutants co-immunoprecipitate with TPR1 and PLCβ2. (A) HEK293 cells were co-transfected with PLCβ2 and the Gα constructs as indicated at the bottom of the blots. Total cell lysates (TCL) from each condition were subjected to immunoprecipitation using either anti-PLCβ2 or anti-Gα16 antiserum followed by protein G-agarose. Well-washed immunoprecipitates were subjected to SDS-PAGE and the proteins of interest were detected using specific antibodies as indicated. TCL were separately run on blots for detecting the corresponding protein expressions. Two separate sets of transfected cells yielded similar results. (B) Similar procedures were applied for detecting the interaction between Flag-tagged TPR1 and the Gα constructs as indicated. TCL were subjected to immunoprecipitation using either anti-Flag agarose gel or anti-Gα16 antiserum followed by protein G-agarose. Two separate sets of transfected cells yielded similar results. Band intensities were quantified and figures on the lanes of QL mutants are the fold increase compared with the band on the lane of their corresponding wild-type counterparts.

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Journal of Molecular Signaling

ISSN: 1750-2187