Mutations in MAPKBP1 Affect JNK2 and/or WDR62 Binding
HEK293 cells were transiently transfected (lipofectamine [ThermoFischer]) with plasmids encoding V5-tagged WT and mutant forms of MAPKBP1 and Myc-tagged WDR62 (A) or HA-tagged JNK2 (B), as indicated. Cells were lysed and immunoprecipitated with the anti-V5 antibody (rabbit polyclonal [Sigma, V8137]). Lysates and immunoprecipitates (IP) were analyzed by western blot as indicated (WB; V5: Mouse monoclonal clone SV5-Pk1 [AbD Serotec MCA-1360], dil 1/5,000; HA: mouse monoclonal, clone 16B12 [Biolegend, 901502], dil 1/5,000; MYC: mouse monoclonal, Ab-2 clone 9E10.3 [Fisher Scientific MS-139-P1], dil 1/5,000).