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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: Exosomal circZNF451 restrains anti-PD1 treatment in lung adenocarcinoma via polarizing macrophages by complexing with TRIM56 and FXR1

Fig. 5

The circZNF451–TRIM56–FXR1 complex regulates the immune phenotype of macrophages. A Silencing of FXR1 in macrophages (PMA stimulated THP-1 cells for 24 h) with the siRNA was verified by western blotting. B The impact of FXR1 knockdown on the M2 phenotype of LPS-stimulated macrophages was detected by flow cytometry. C Concentrations of IFN-γ and GZMB in the supernatant of CD8+ T cells cocultured with the supernatant from macrophage-siFXR1 for 48 h were detected by ELISA. D After LPS stimulation, concentrations of GM-CSF, CXCL1, IL-1β, IL-1Ra, and IL-10 in the supernatant of macrophage-siFXR1 were measured by ELISA. E The impact of TRIM56 knockdown on the M2 phenotype of LPS-stimulated macrophages after coculturing with H1395-circZNF451 for 48 h was analyzed by flow cytometry. F The M2 phenotype markers Arg1 and IL-10 were measured by qRT-PCR after silencing TRIM56 in LPS-stimulated macrophages cocultured with H1395-circZNF451 for 48 h. G Concentrations of IFN-γ and GZMB in the supernatant of CD8+ T cells cocultured with the supernatant from the macrophage-siTRIM56/H1395-circZNF451 for 48 h were detected by ELISA. H Concentrations of GM-CSF, CXCL1, IL-1β, IL-1Ra, and IL-10 in the supernatants of H1395-circZNF451/macrophage-siTRIM56 stimulated by LPS were measured by ELISA. I After LPS stimulation, concentrations of GM-CSF, CXCL1, IL-1β, IL-1Ra, and IL-10 in the supernatants of macrophage-TRIM56mutant/A549 cells were measured by ELISA. B-D and I used the two-tailed, Student’s t test. EH were analyzed by one-way ANOVA test after adjusting for multiple comparisons. All experiments with statistical analysis have been repeated for at least three times

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