Fig. 5

APG-2575 enhanced M1 polarization by upregulating NLRP3 expression. A Quantification of CD86 and CD206 expression in IL-4-activated Bcl-2-knockout or Bcl-2--overexpressing RAW264.7 cells treated with APG-2575 or control. B–F KEGG pathway analysis and GSEA using genes differentially expressed between IL-4-activated RAW264.7 cells with the control or APG-2575 treatment. G–L Scatter plot showing the results of Pearson correlation analysis between the estimated M1 macrophage infiltration score and NLRP3 gene expression level in various cancer types. M Representative immunofluorescence staining of NLRP3, ASC, and DAPI in IL-4-activated BMDMs after treatment with APG-2575 in combination with JSH-23 or INF39. Scale bar, 20 μm. N Western blot analysis of NOS2, NLRP3, caspase-1, Arg-1, and IL-1β in IL-4-activated BMDMs cultured with APG-2575 in the presence or absence of INF39. O, P The mRNA expression levels of M1/M2-related markers in IL-4-activated BMDMs treated with APG-2575 in the presence or absence of INF39. Q Quantification of CD86, MHC-II and CD206 expression in IL-4-activated BMDMs treated with APG-2575 in the presence or absence of INF39. R IL-4-activated CD14+ monocyte-derived macrophages treated with or without APG-2575. The TNF-α concentration in the supernatants was measured by a CBA, and the IL-10 concentration in the supernatants was measured by ELISA. S, T Western blot analysis of NOS2, NLRP3, caspase-1, Arg-1, and IL-1β in IL-4-activated BMDMs with stable Nlrp3 knockdown and overexpression and treated with APG-2575 or control. LUAD, lung adenocarcinoma, LUSC, lung squamous cell carcinoma, OV, ovarian serous cystadenocarcinoma, COAD, colon adenocarcinoma; BLCA, bladder urothelial carcinoma, ESCA, esophageal carcinoma