Application
Wild-type (+/+) and Gab1-/- (-/-) cells were heat shocked (HS) at 42掳C for 1 h or irradiated with UV-B light (400 J/m2) and then incubated at 37? for 1 h. Cell lysates were prepared and subjected to immunoprecipitation (IP) with an anti-JNK2 antibody. The immunoprecipitates were resolved by SDS- PAGE and immunoblotted (IB) with anti-Gab1 antibody. Data courtesy of Dr. GS Feng at The Burnham Institute.
Application
Rescue of the JNK pathway by expression of wild-type Gab1 in Gab1-/- cells. Gab1-/- cells were transiently transfected with 1, 2, or 4 of the expression construct for human Gab1 cDNA, using the GenePORTER 2 transfection reagent (Gene Therapy Systems Inc.). After incubation, the cells were irradiated with UV-B light at 400 J/m2 or left untreated. Exogenous expression of Gab1 in transfected cells was confirmed by anti-Gab1 immunoblot analysis with wild-type cells as positive control. +/+, wild-type cells; -/-, Gab1-/- cells. (Mol. Cell. Biol. 2004 Feb 15;24(4):1531-1539)