Fig. 5

Migration, proliferation and angiogenesis ability of HUVECs co-cultured with four groups of Mφs (siNC, siNC + succinate, siSUCNR1 and siSUCNR1 + succinate). A, B Images were measured at 0 h, 6 h, 12 h and 24 h in the scratch wound healing test. The cell migration rate was used to indicate migratory ability as described in the article. Scale bar: 200 μm (n = 3). C, D In the Transwell assay, ImageJ software was used to calculate the stained cells in the four groups. Scale bar: 200 μm (n = 3). E, G EdU assays revealed the proliferation of HUVECs, and the percentage of EdU-positive cells in the four groups was calculated. Scale bar: 200 μm (n = 3). F Cell viability was tested by CCK8 assay, and the OD at 450 nm was recorded to calculate cell viability at 24 h and 48 h (n = 3). J Tubular formation of four groups was recorded after being cultured on Matrigel for 6 h. Scale bar: 200 μm. The number of branches (H), junctions (I), total tube length (K) and increase of tube formation (%) (L) were qualified by ImageJ software (n = 3). M–O Relative protein levels of CD31 and VEGF-A in the four groups (n = 3). P Immunofluorescence staining for DAPI (blue), VEGF-A (green) and CD31 (red) in HUVECs. Scale bar: 50 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001