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Fig. 10 | Journal of Nanobiotechnology

Fig. 10

From: Doxorubicin-polyglycerol-nanodiamond conjugate is a cytostatic agent that evades chemoresistance and reverses cancer-induced immunosuppression in triple-negative breast cancer

Fig. 10

M2–M1 phenotypic shift of tumor-associated macrophages induced by Nano-DOX-treated 4T1 cells. a, b Immunostaining of CD86 (an M1 marker) in type-2 macrophages (M2) assayed by FACS. c Expression of GBP5 (an M1 marker) in M2 assayed by western blotting. d mRNA expression of iNOS (an M1 marker) in M2 assayed by RT-PCR. e, f mRNA expression of CD206 and TGF-β (M2 markers), respectively in M2 assayed by RT-PCR. g Immunohistochemical staining of CD80, CD86, GBP5, MHC-II (M1 markers), and CD206 (an M2 marker) in 4T1 tumor xenograts at the end of 3-week treatment of Nano-DOX or DOX. In the ex vivo cell experiments, type-1 and type-2 macrophages (M1 and M2) were activated from mouse bone marrow-derived macrophages according to published protocols. M2 were treated with Nano-DOX (2 μg/mL) or culture medium conditioned by Nano-DOX (2 μg/mL)-treated 4T1 cells (ND-CM). Duration of Nano-DOX or Nano-DOX-CM treatment was 24 h. In FACS analysis, geometric means were used to quantify fluorescence intensity. Values were mean ± SD (n = 3, *p < 0.05, **p < 0.01)

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