Figure 2
CECR1 promotes the proangiogenic paracrine action of M2 macrophages. (a) Diagram showing the experimental setup of testing paracrine angiogenic activation of vascular cells in 3D co-cultures by THP1 macrophages. (b) Low and high-magnification fluorescent images of neo-vessel formation by HUVECs (GFP-marked) and human-derived pericytes (dsRed-marked) without THP1 macrophage stimulation. (c) Low- and high-magnification fluorescent images of neo-vessel formation by HUVECs (GFP-marked) and human-derived pericytes (dsRed-marked) with stimulation of non-treated (control), and sisham- or siCECR1-treated THP1 macrophages. (d) Low- and high-magnification fluorescent images of neo-vessel formation by HUVECs (GFP-marked) and human-derived pericytes (dsRed-marked) with stimulation of non-treated (control), and sisham- or siCECR1-treated THP1 macrophages with U87 stimulation (scale bar, 100 μm for b–d). (e) Upper image: western blot of CECR1 protein and β-actin loading control in THP1 macrophages. Blot represents results from three observations. Lower graph: QPCR results of CECR1 mRNA levels normalized to housekeeping genes in non-treated (control), and sisham- or siCECR1-treated THP1 macrophages, without and with U87 stimulation. *P<0.05; **P<0.01. (f–h) Quantified results of the co-culture experiment. Number of junctions, tubules and total tubule length data are shown. *P<0.05; **P<0.01. (MФ (-) versus MФ-control, U87 MФ-control versus MФ-control, MФ-sisham versus MФ-siCECR1, U87 MФ-sisham versus U87 MФ-siCECR1). Representative graphs were taken from at least three experiments. Six wells were analyzed in each experiment.
