Fig. 2

P4HA1 overexpression promotes angiogenesis in vitro and in vivo. A Validation of Ad-P4HA1 overexpression efficiency in HUVECs (n = 3). B–D After HUVECs were infected with Ad-NC and Ad-P4HA1 for 24 h, B EdU incorporation assay (magnification: ×100, scale bar: 100 μm), C tube formation assay (magnification: ×100, scale bar: 100 μm), and D transwell migration assay (magnification: ×400, scale bar: 50 μm) were performed (n = 3). E In vivo, P4HA1 overexpression was achieved by injecting purified adenoviruse into the gastrocnemius muscle on the first and seventh day after surgery. Overexpression efficiency of P4HA1 in gastrocnemius muscle of the HLI model (14 d) was detected and analyzed (n = 5). F Representative Doppler images of blood flow recovery after the HLI model with P4HA1 overexpression. G Statistical analyses of blood flow recovery in HLI model with P4HA1 overexpression (n = 5). H Immunofluorescence staining of CD31 (green) in the gastrocnemius muscle from the HLI model (14 d) with P4HA1 overexpression. magnification: ×400, scale bar: 50 μm. The fluorescence intensity was statically analyzed (n = 5). Data in A–E, H were analyzed using the Student’s t-test. Data in G were analyzed by two-way ANOVA with Bonferroni post hoc test. *p < 0.05. NC negative control, HLI hindlimb ischemia