FIGURE 6.

Ppp2r2d and Ampkα co-localize in A7r5 cells. A7r5 cells were seeded on poly-l-lysine treated coverslips and cultured in DMEM with 10% fetal bovine serum overnight. Cells were fixed with 4% paraformaldehyde in PBS followed by a permeabilization step with 0.25% Triton X-100. The cells were blocked in 10% donkey serum in 0.05% Triton X-100, PBS and then treated with a 1:50 dilution of primary antibodies (PPP2R2D (Abcam)) and AMPKα1 antibodies (Santa Cruz Biotechnology). The coverslips were incubated with secondary antibodies (Cy3-conjugated rabbit antibodies and DyLight 488-conjugated goat antibodies). Coverslips were mounted in Fluoromount-G, imaged by a Leica DMI6000B microscope, and analyzed by ImageJ software.