Fig. 4.

Augmentation of ADAM12 expression in Balb/c improved perfusion recovery, whereas knockdown of ADAM12 expression in C57Bl/6 by small hairpin (sh)RNA impaired perfusion recovery following experimental PAD. A: higher ADAM12 mRNA is expressed in hindlimbs that received ADAM12 cDNA compared with those that received control plasmid, 10 days post-treatment (n = 5/group, *P < 0.05). B: ADAM12 Western showing higher expression in muscle lysates from mouse hindlimbs that received ADAM12 cDNA (top); bottom: quantification of bands in top (n = 3/group, *P = 0.04). C: representative laser Doppler perfusion imaging (LDPI) image showing increased perfusion in Balb/c mice that received ADAM12 cDNA. D: LDPI showing perfusion recovery after HLI. y-Axis shows extent of perfusion in the ischemic limb relative to the nonischemic limb; x-axis shows days and weeks following experimental PAD (n = 7–10/group, *P < 0.05). E: anti-CD31 staining of sections of ischemic hindlimbs, 5 wk following experimental PAD, showed a higher number of capillaries/muscle fiber (capillary density) in Balb/c mice that received ADAM12 cDNA (control n = 5, ADAM12 plasmid n = 7, *P = 0.05). F: ischemic hindlimbs from mice treated with shRNA targeting mouse ADAM12 showed decreased ADAM12 mRNA expression compared with hindlimbs treated with control plasmid (n = 6, ***P < 0.01), 10 days postinjection. G: shADAM12-treated hindlimbs show no change in expression of 2 other ADAM isoforms (ADAM15 and ADAM17, n = 6, NS = P > 0.05). H: representative LDPI image showing impaired perfusion in C57Bl/6 mice that received plasmid containing mouse ADAM12 shRNAs. I: y-axis shows extent of perfusion in the ischemic limb relative to the nonischemic limb; x-axis shows days and weeks following experimental PAD (n = 13/group, *P < 0.05). J: perfusion in nonischemic, shADAM12-treated hindlimbs is not different from perfusion in control, nonischemic hindlimbs (NS = P > 0.05, n = 5 for shADAM12, and n = 6 for control).