FIG 3.

GA stimulates an association between MLK3 and Hsp70. (A) HEK293 cells were transfected with the indicated combinations of FLAG-MLK3 and GFP-Hsp70 WT plasmids. The cells were treated with DMSO or GA (5 μM) for 3 h. FLAG-MLK3 immunoprecipitates were immunoblotted with anti-GFP and anti-FLAG antibodies. GFP-Hsp70 immunoprecipitates were also immunoblotted with anti-GFP antibody. Whole-cell extracts were immunoblotted with anti-FLAG, anti-GFP, and anti-β-actin antibodies. (B) SKOV3 cells were treated with GA (10 μM) for the indicated time periods, and endogenous MLK3 immunoprecipitates were immunoblotted with anti-Hsp90 and anti-Hsp70 antibodies. Whole-cell extracts were also immunoblotted with anti-MLK3, anti-Hsp70, anti-Hsp90, and anti-β-actin antibodies, as indicated.