Fig. 2

VNP-shCCDC25 stimulates NEs and macrophage activation and infect cells in vitro. A The expression of N1 polarization markers of NEs after VNP-shCCDC25 treatment was analyzed via qPCR. B The expression of N2 polarization markers of NEs after VNP-shCCDC25 treatment was analyzed via qPCR. C The expression of M1 polarization markers of RAW264.7 cells after VNP-shCCDC25 treatment was analyzed via qPCR. D The expression of M2 polarization markers of RAW264.7 cells after VNP-shCCDC25 treatment was analyzed via qPCR. E Schematic diagram of RAW264.7 cell medium stimulated by VNP-shCCDC25 promotes apoptosis of B16F10 cells. After VNP-shCCDC25 was co-cultured with RAW264.7 cells for 2 h, the medium was changed to fresh medium containing gentamycin, and the incubation was continued for 6 h. After 6 h, the RAW264.7 cell medium was collected and co-incubated with B16F10 cells for 16 h. Finally, B16F10 cells were collected for the apoptosis assay. F The apoptosis levels of B16F10 cells after incubated for 16 h with RAW264.7 cell medium, which was stimulated with VNP-NC, VNP-shCCDC25, or not (n = 3). G The apoptosis levels of B16F10 cells after incubated with VNP-NC, VNP-shCCDC25, or not for 16 h (n = 3). H The fluorescent pictures of B16F10 cells incubated with VNP-RFP (MOI = 100:1) or not; actin (green), DAPI (blue), VNP-RFP (red). Scale bars: 25 μm. I The titer of bacterium colonized in the B16F10 cells after co-incubated with VNP-NC or VNP-shCCDC25 (n = 3) for 16 h. J The expression of CCDC25 in B16F10 cells after incubated with VNP-shCCDC25 was analyzed via qPCR. Data are shown as the mean ± SD. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns: no significance