Fig. 8.

AMPK inhibition reduces CSE phosphorylation, per-polysulfide and ischemic blood flow. Panel A Representative blots of ischemic gastrocnemius muscle tissues from mice subject to the femoral artery ligation (FAL) 0hrs, 3hrs, 24hrs and 5 days probed for pCSES346, total CSE, pAMPK, AMPK and GAPDH. Panel B Graphic representation of the densitometry quantification depicted in Panel A for pCSE346 and p-AMPK protein expression. Panel C Representative western blots were performed from non-ischemic (NI), ischemic (Isch), and ischemic AMPK-I (Isch + AMPK-I) skeletal muscle (SkM) tissues collected at day 5 post femoral artery ligation. Panel D Graphic representation of the densitometry quantification of pCSE346 and p-AMPK protein expression. Panel E Representative angiogram images of hindlimbs showing blood flow (blue color = lowest flow; red color = highest flow) from saline treated post-ligation at day 5 (upper panel; Sal-D5) and AMPK inhibitor treated post-ligation at day 5 (lower panel; AMPK-I-D5). Panel F Graphic representation of the densitometry quantification depicted in Panel E for ischemic limb blood flow. Panel G Plasma sulfide levels, including free/acid labile pools (F/Al) and bound sulfide of pre-FAL, saline (sal) or AMPK-I treated mice. In all densiometric analysis of western blots, p-AMPK and p-CSE346 are normalized to total AMPK or total CSE that are normalized to GAPDH. All the data were representative of at least n = 5. ****P < 0.0001; **P < 0.003; *P < 0.01. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)