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. Author manuscript; available in PMC: 2015 Nov 15.
Published in final edited form as: Biochem J. 2014 Nov 15;464(1):99–108. doi: 10.1042/BJ20140420

Figure 3. Paxillin associates with MEKK2 independent of kinase activity.

Figure 3

(A) Anti-MEKK2 immunoblot of MDA-MB 231 cells (left lane)with stable MEKK2 knockdown (center lane)and expression of “added back” shRNA-resistant 6xHis-tagged MEKK2 (right lane). (B) Immunoblot analysis shows the association between MEKK2 and paxillin in MDA-MB 231 breast tumor cells. Endogenous paxillin was co-purified with MEKK2 from MDA-MB 231 cells with stable expression of 6xHis-tagged MEKK2 (top panel) by Ni-NTA bead affinity column pull down. Also shown are immunoblots detecting c-Raf (middle panel, negative control) and MEKK2 (bottom panel). (C) HA-tagged wild type MEKK2 or kinase inactive mutant (K385M) MEKK2 co-immunoprecipitates with FLAG-tagged paxillin in transfected 293T cells (upper two panels). Immunoblots of lysates show loading in the lower two panels. The results are representative of at least three independent experiments.