Fig. 3

Cyclin G2 in macrophages suppresses tumors by inhibiting tumor blood vessels after IFN-γ treatment. A CD31 immunohistochemical staining of the LLC tumors isolated from mice in the WT and Ccng2^−/− groups, representative images are shown. 10× Scale bar = 200 μm; 40× Scale bar = 50 μm. B Graph showing the number of blood vessels in each field (n = 5). Data were analyzed using the unpaired Student’s t-test. Data are presented as the mean ± SEM. C, D Tube formation of SVEC4–10 cells treated with conditioned medium from BMDMs isolated from WT and Ccng2^−/− C57BL/6 mice. Scale bar = 500 μm (representing 3 independent experiments). E Western blot showing successful knockdown of cyclin G2 in THP-1 cells. GAPDH was used as a loading control. F RT-qPCR was used to measure CCNG2 mRNA expression levels in THP-1 stable cell lines (Nonsense, shcyclin G2#1, and shcyclin G2#2). β-actin was used as an internal control. G Western blotting demonstrated successful cyclin G2 overexpression in THP-1 cells. β-tubulin was used as a loading control. H Measurement of CCNG2 mRNA expression levels in THP-1 stable cell lines (Vector and Flag-cyclin G2) by RT-qPCR. β-actin was used as an internal control (representing 2 independent experiments). I Tube formation by HUVECs treated with conditioned medium from THP-1 stable cell lines (Nonsense, shcyclin G2#1, and shcyclin G2#2) cells. Scale bar = 200 μm (representing 3 independent experiments). J Tube formation by HUVECs treated with conditioned medium from THP-1 stable cell lines (Vector and Flag-cyclin G2) cells. Scale bar = 200 μm (representing 3 independent experiments). Data in D, F, H, I, and J were analyzed with the unpaired Student’s t-test. Data are presented as the mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant