Fig. 2

Overexpression of KPNA2 promoted angiogenesis in vitro and in vivo under hypoxia. A HUVEC were infected with Ad-KPNA2 to overexpress KPNA2. After 48 h of infection, tube formation assays were carried out under normoxia and under hypoxia, and pictures were taken 6 h later. Scale bar: 200 μm. The total lengths of the tube formation assay were analyzed statistically. Each experiment was repeated three times. B HUVEC were infected with Ad-KPNA2 to overexpress KPNA2. After 48 h of infection, Transwell assays were performed under hypoxia, and pictures were taken 6 h later. Scale bar: 200 μm. For each experiment, 3 fields of view were randomly selected for cell counting, and the average value was taken for statistical analysis. Each experiment was repeated three times. C HUVEC were infected with Ad-KPNA2 to overexpress KPNA2. After 48 h of infection, EdU staining was carried out under hypoxia, and pictures were taken 6 h later. Scale bar: 200 μm (EDU: red, DAPI: blue). EDU/DAPI were analyzed statistically. Each experiment was repeated three times. D Changes in blood perfusion in the ischemic hindlimb of model mice after overexpression of KPNA2 before and after surgery and at 3 days, 7 days, and 14 days. n = 5. E CD31 immunofluorescence (green) analysis of the gastrocnemius muscle of the ischemic mouse hindlimb at 14 days after KPNA2 overexpression. Scale bar 100 μm. n = 5. F Statistical analysis of blood flow changes in the ischemic hindlimb of model mice after overexpression of KPNA2 before and after surgery and at 3, 7, and 14 days. n = 5. G Vessel density statistics of CD31 immunofluorescence of the gastrocnemius muscle of the ischemic mouse hindlimb at 14 days after KPNA2 overexpression. n = 5. H KPNA2 expression of the gastrocnemius muscle of the ischemic mouse hindlimb at 14 days after KPNA2 overexpression detected by Western Blot. n = 5. * p < 0.05. NonHLI non-hindlimb ischemia, HLI hindlimb ischemia