FIGURE 7: FLCN is associated with phospho-AMPK and LKBI mobilization during shear stress. HK2 cells were transfected with control siRNA (siCTRL) or with siRNA targeting FLCN (siFLCN). 72 h later, they were subjected (shear) or not (static) to fluid flow for the indicated times. (A) Phospho-AMPK (Thr172), total AMPK, FLCN and actin levels were analyzed by western blot and quantified (B) in the indicated conditions (A). (C, D) Cells prone to fluid flow (shear 4D) or not (static 4D) were fixed with methanol, labeled with DAPI, immunostained for ARL13B, phospho-AMPK (C), LKB1 (D), ARL13B, and then analyzed by fluorescence microscopy. Arrowheads indicate presence of phospho-AMPK (P-AMPK) or LKB1 at primary cilium. Scale bars in (C) and (D) = 10μm.