Fig. 4.

DUSP13A has no effect on Akt1 activity and ASK1 oligomerization. a HEK 293 cells were transfected with 0.5, 1, or 2 μg of FLAG-DUSP13A expression plasmid. After transfection, cells were lysed with PTP lysis buffer. Cell lysates were subjected to immunoblotting using anti-Akt1 and anti-phospho-Akt1 (Ser-473) antibodies. b HEK 293 cells were separately transfected with 1 μg of FLAG-ASK1 or HA-ASK1 plasmid. After 48 h of transfection, cells were lysed in lysis buffer. Both cell lysates were mixed and incubated in the presence of DUSP13A WT or mutant. FLAG-ASK1 was immunoprecipitated with anti-FLAG M2 affinity agarose. Pulled-down ASK1 complexes were immunoblotted with an anti-HA antibody