Figure 9: AMPK is recruited onto F-actin by directly binding the LIM domain of Pdlim5.
From: Augmented AMPK activity inhibits cell migration by phosphorylating the novel substrate Pdlim5
(a) HEK293T cells were co-transfected with V5-tagged AMPKα and FLAG-tagged Pdlim5 (WT, ΔPDZ or ΔLIM domain). TCLs were immunoprecipitated by FLAG or V5 and then immunoblotted with the indicated antibodies. (b) F-actin binding assay of AMPK. AMPK was mixed with a fixed amount of F-actin in the presence or absence of α-actinin and GST-Pdlim5, incubated for 1 h at 24 °C and then centrifuged at 150,000 g for 1.5 h at 24 °C, to pellet the F-actin polymer and associated proteins. A sample of the pellet (P) and supernatant (S) were analysed by immunoblotting for the indicated antibodies.
