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. 2020 Apr 14;11:1833. doi: 10.1038/s41467-020-15290-0

Fig. 4. NHWD-870 reduced the proliferation of TAMs in vivo.

Fig. 4

a, b NHWD-870 treatment reduced the number of TAMs in subcutaneously implanted H526 and A2780 tumors. Shown are immunofluorescent staining of CD68 (macrophage marker, red) and E-Cadherin (cancer cells, green) (a) and quantification of CD68 positive cells (b) in vehicle, NHWD-870 treated H526 (n = 6) and A2780 (n = 5) tumors from Fig. 3a, b, respectively. Scale bar is 100 μm. Data are presented as mean ± SEM from different tumors. c, d NHWD-870 treatment reduced the proliferation TAMs in subcutaneously implanted A2780 tumors. Shown are representative immunofluorescent staining of Ki67 and CD68 (c) and quantification of percentage of Ki67+ cells in CD68+ cells (d). Scale bar is 30 μm. Data are presented as mean ± SEM of five different tumors. e Schematics of the animal experiments using an orthotopic mouse OC model, established by intraperitoneally injecting human A2780 cells into female recipient nude mice. Mice were then either untreated (Ctrl) or treated with NHWD-870 (PO, QD, 3 mg/kg) for 45 days. Half of the recipient mice received A2780 cells plus TAMs isolated from tumors of OC-bearing donor mice. f, g Tumor weights (f) and ascitic fluid volumes (g) at day 45 for mice in (e). Data are presented as mean ± SEM from five different mice. h–k NHWD-870 treatment decreased proliferation of the both tumor cells and TAMs. Shown are representative immunofluorescent staining of Ki67 and CD68 (h) and quantification of Ki67+CD68− (i) or Ki67+CD68+ (j), or CD68+ cells (k). Scale bar is 50 μm. Data are presented as mean ± SEM from five different mice. p values were calculated using two-tailed, unpaired t tests in this figure. *p < 0.05; **p < 0.01; ***p < 0.001. Source data are provided as a Source Data file.