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. 2025 Jul 16;17(14):2365. doi: 10.3390/cancers17142365

Figure 3.

Figure 3

Co-culture of HCC cells with macrophage assays to verify the correlation between MICA and macrophage polarization. (a) Western blotting assay confirms increased MICA level in Huh-7 cells with stably over-expressed MICA compared to the control vector (NC); the statistically quantificational results are shown on the right (n = 4). The uncropped blots are shown in Supplementary File S1. (b) The CD86 mRNA expression is shown in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h (n = 9). (c) The CD206 mRNA expression is shown in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h (n = 8). (d) The TNF-α mRNA expression is shown in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h (n = 8). (e) The IL-10 mRNA expression is shown in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h (n = 6). (f) The TNF-α levels in supernatants from cultured macrophages were measured by ELISA (n = 5). (g) The IL-10 levels in supernatants from cultured macrophages were measured by ELISA (n = 9). (h) Representative immunofluorescence staining for CD86 (n = 4) and CD206 (n = 5) in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h and statistically quantificational results. (i) Representative immunofluorescence staining for TNF-α (n = 3) and IL-10 (n = 6) in macrophages co-cultured with MICA+ Huh-7 cells or NC cells for 24 h or 72 h and statistically quantificational results. All images were taken at 400× magnification. p-value significance codes: * p < 0.05, ** p < 0.01, and *** p < 0.001.