Fig. 4
From: PIM kinases inhibit AMPK activation and promote tumorigenicity by phosphorylating LKB1
Ser334 is a PIM target site in LKB1 and is essential for increased AMPK phosphorylation in response to PIM inhibition. a PC3 and MCF7 cells were transiently transfected with the FLAG-LKB1 plasmid. After 24 h, cells were treated with either DMSO or 10 μM DHPCC9 for another 24 h. 10% of cell lysates were stained with FLAG antibody (Input), while the rest was immunoprecipitated with FLAG M2 affinity gel and stained with PAS antibody (IP:FLAG). Shown are representative images as well as graphs with relative differences in phosphorylation levels of LKB1 as compared to the DMSO-treated control samples (average values ± SD, n = 3). b MCF7 WT and TKO cells were transiently transfected with WT or S334A FLAG-LKB1 plasmids. After 48 h, relative phosphorylation levels of LKB1 were determined (average values ± SD, n = 3). c LKB1 KO derivatives of PC3 and MCF7 cells were transiently transfected with FLAG or FLAG-LKB1 (WT or S334A) plasmids. After 24 h, cells were treated with either DMSO or 10 μM DHPCC9 for another 24 h before Western blotting with pAMPK (Thr172), AMPK and LKB1 antibodies. Shown are representative images as well as graphs with relative phosphorylation levels of AMPK as compared to DMSO-treated control samples (average values ± SD, n = 3)