Phospho-CDK5 (Tyr239) Antibody - #AF3628

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 

Figure 2 Neutrophils infiltration and NETs formation in mice brain tissue. (A) Representative immunofluorescence staining of Ly6G-positive neutrophils (red) and H3cit -positive(green) and CD31-labeled blood vessels (white) in the mice brain tissue sections post-CCT 3 days. Nuclei were stained with DAPI (blue). (B) Representative Western blot bands of MPO, PAD4, H3Cit, and H3 and statistical analysis expression level of MPO (C), PAD4 (D), and H3Cit (E) in mice brain tissue (n=6). Data are represented as mean ± SD. *P < 0.05, **P < 0.01.

Figure 6. Inhibition of PYGB suppresses HCC growth. (A) HCC cell viability analysis of CP91149 treatment. (B) HCC cell viability analysis of CP91149 treatment combined with sorafenib. (C) Tumor growth curve of HCC under vehicle (n=6), CP91149 (n=6), sorafenib (n=6), or combination treatment (n=6). (D) HCC tumor weight. (E) HCC tumor pictures. (F) H&E (scale bar, 200 µm) staining of tumor tissues and immunohistochemical analysis of Ki67 (scale bar, 100 µm) and CD31 (scale bar, 50 µm). (G) Quantification of Ki67 positive cells. (H) Quantification of blood vessel density. *P