AMP-activated protein kinase (AMPK) plays a critical role in the stimulation of glucose transport in response to hypoxia and inhibition of oxidative phosphorylation. Here we examine the signaling pathway(s) mediating the glucose transport response following activation of AMPK. Using mouse fibroblasts of AMPK-wild-type and AMPK-knockout, we documented that the expression of AMPK is essential for the glucose transport response to both azide and AICAR. In Clone 9 cells, the stimulation of glucose transport by a combination of azide and AICAR was not additive, while there was an additive increase in the abundance of P-AMPK. In Clone 9 cells, AMPK-wild-type fibroblasts, and in H9c2 heart cells, azide or hypoxia selectively increased P-ERK1/2, while, in contrast, AICAR selectively stimulated P-p38; phosphorylation of Jun-activated kinase (JNK) was unaffected. Azide's effect on P-ERK1/2 abundance and glucose transport in Clone 9 cells was partially abolished by the MEK1/2 inhibitor, U0126. SB203580, an inhibitor of p38, prevented the phosphorylation of p38 and the glucose transport response to AICAR and, unexpectedly, to azide. Hypoxia, azide, and AICAR all led to increased phosphorylation of AS160 in Clone 9 cells. Employing siRNA directed against AS160 did not inhibit the glucose transport response to azide or AICAR, while the content of P-AS160 was reduced by ~80%. Finally, we found no evidence for co-immunoprecipitation of Glut1 and P-AS160. We conclude that while azide, hypoxia, and AICAR all activate AMPK, the downstream signaling pathways are distinct with azide and hypoxia stimulating ERK1/2 and AICAR stimulating the p38 pathway.