Figure 3.

Effect of glucose starvation on various signaling pathways in HEK293T cells. HEK293T cells were maintained in serum-free DMEM containing 5 mm glucose for ∼6 h and then treated with serum-free DMEM containing 0 or 5 mm glucose for 10 min. Following treatment, whole-cell lysates were prepared as described under “Experimental procedures.” Samples were subjected to SDS-PAGE, and Western blotting was performed using antibodies directed against the phosphorylated substrates of various signaling pathways, including PI3K (P-Akt), ERK (P-ERK), PKA (P-VASP), and AMPK (P-ACC). Total levels of the substrates were also detected using the corresponding antibodies. Representative images are shown. n = 3.