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. 2008 Oct 24;105(44):17085–17090. doi: 10.1073/pnas.0802701105

Fig. 4.

Fig. 4.

Interaction of CARMA3 with β-arrestins. (A) HEK293T cells were transfected with expression vectors encoding HA-tagged CARMA3, FLAG-tagged β-arrestin 1, or FLAG-tagged β-arrestin 2 at different combinations. Twenty hours after transfection, cell lysates were subjected to immunoprecipitation (IP) with anti-FLAG antibody-conjugated beads, and immunoprecipitated proteins and cell lysates were subjected to SDS/PAGE and analyzed by immunoblotting with the indicated antibodies. (B) Wild-type or β-arrestin 2 KO MEF cells (15-cm plate, 80% confluent) were stimulated with or without LPA for different periods of time and then scraped off and lysed. The resulting lysates were subjected to immunoprecipitation with CARMA3 antibody-conjugated beads. The obtained immunocomplexes were subjected to SDS/PAGE and Western blotting with β-arrestin 1 and β-arrestin 2 antibodies or CARMA3 antibodies as indicated. For detection of coimmunoprecipitated β-arrestins, rabbit IgG TrueBlot was used as secondary antibody.