Fig. 5

B.adolescentis regulated DCN⁺ macrophages through TLR2/YAP axis. A, B BMDMs and THP-1 cells were incubated with B.adolescentis or vehicle (PBS) for 24 h. Protein levels of TLR2 and YAP were tested by Western blot. C Immunofluorescence assay of YAP distribution in BMDMs. BMDMs were stained with specific antibody against YAP (green), and the nuclei were counterstained with DAPI (blue). Scale bar, 10 μm. D, E Nuclear and cytoplasmic separation assay was performed in B.adolescentis-treated BMDMs and THP-1 cells, then the protein level of YAP in the nuclear and cytoplasm was detected by Western blot. F, G BMDMs and THP-1 cells were incubated with B.adolescentis or vehicle (PBS) for 24 h with or without 1 μM verteporfin. Protein levels of TLR2, YAP and DCN were tested by Western blot. H, I YAP was overexpressed in BMDMs and THP-1 cells, and protein levels of YAP and DCN were tested by Western blot. J-L HCT116 cells were injected into BALB/c nude mice combined with THP-1 cells pretreated with B.adolescentis or vehicle (PBS) for 24 h (n = 5 per group). From the beginning of tumor inoculation until sacrifice, 50 mg/kg verteporfin or vehicle (5% DMSO) were injected intraperitoneally to mice every day. Tumor volume was recorded after 6 days. The independent experiment was repeated three times. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, **** P < 0.0001; ANOVA test (K, L)