Fig. 7
Blocking VEGF-A signalling inhibits homing of proangiogenic monocytes to tumours. a Analysis of Cx3cl1 protein expression in xenografts of SKBR7, DLD1 and HCT116 tumours. b Quantification of Cx3cl1 band intensity in a after normalisation with same sample Actin level. c Scheme of the study of VEGF-A implication in human monocyte recruitment in DLD1 xenograft. DLD1 xenograft-bearing mice were treated either with the mixture of DC101 and bevacizumab (D/B mix) or control IgG for 24 h before adoptive transfer of human pan monocytes. The mice were perfused with PBS-EDTA to wash out circulating and non-extravasated leucocytes before tumour collect and analysis. d Analysis of Cx3cl1 protein level in DLD1 tumour after 24 h of D/B mix treatment by western blotting. e Quantification of Cx3cl1 band intensity in d after normalisation with Pecam1 level in same samples. f Expression level of Cx3cl1 in tumour vasculature by immunofluorescence of DLD1 xenografts. Scale bar = 80 µm. g Quantification of Cx3cl1 in blood vessels. Data are presented as scatter dot plots with medians. Each dot represent the mean intensity of Cx3cl1 per µm2 blood vessels from 10 sections of the same tumour. n = 5 tumours per group were used for this quantification. **p < 0.005 in Mann–Whitney test. h FACS analysis of the effect of D/B mix on human proangiogenic monocyte recruitment to DLD1 xenograft. i Effect of blocking VEGF-A signalling on extravasation of proangiogenic monocytes from different donors. The data presented as aligned dot plot with medians. n = 6 tumours per group in independent adoptive transfer experiments with monocytes from three donors (DonorA, DonorB and DonorC). *p < 0.05, **p < 0.005, ***p < 0.001 in multiple t-test
