Fig. 4
ADNVs are preferentially taken by TAMs and reprogram their phenotypes. A The simplified experimental scheme for B-D. C57BL/6 J mice were implanted with LLC cells for 7 d and then inoculated with clodronate liposomes (CL) or PBS-liposomes (PL) every 4 days to deplete TAMs. The mice were simultaneously administrated with ADNVs, and sacrificed at day 21 post implantation. B Gross photos of tumors at the end of experiments. C Tumor growth profiles. **p < 0.01 (Two-way ANOVA and Bonferroni post-tests). D Tumor weights at the end of the experiment. **p < 0.01 (One-way ANOVA and Tukey’s significant difference post hoc test). E Schematic outline of adoptive transfer of macrophages for F–H. M2 macrophages were prepared from BMDMs by stimulated with IL-4, and then treated with or without ADNVs (20 μg/mL). M1 macrophages, prepared by stimulation of IFN-ɣ, were used as a positive control. After that, the cells were adoptively transferred to tumor-bearing mice at day 14 post LLCs implantation. Mice were sacrificed at day 21. F Gross photos of tumors at the end of experiments. G Tumor growth profiles. *p < 0.05, ***p < 0.001 (Two-way ANOVA and Bonferroni post-tests). H Tumor weights at the end of the experiment. *p < 0.05, ***p < 0.001 (One-way ANOVA and Tukey’s significant difference post hoc test). I, J ADNVs were stained with Dil and injected into tumor-bearing mice (25 mg/kg, i.p.). Flow cytometry of Dilpos cells in macrophages and other cell populations in tumors (I). Immunofluorescence (IF) staining of BMDMs and other cell lines taking Dil-labelled ADNVs upon incubation. Scale bar = 200 μm. Nuclear: DAPI (J). (K-M) BMDMs were incubated with ADNVs or vehicles for 24 h. Quantification of M1 (CD86+) and M2 (CD206+) population by flow cytometry (K); ELISA assay of TNF-α and IL-6 levels (L); Flow cytometry of ROSpos macrophages and quantification of the mean fluorescence intensity (MFI) (M). *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t-test). The results are from one of two or three independent experiments. Shown are representative images, and the data are presented as means ± SEM