Journal: Cell Reports Medicine
Article Title: Macrophage efferocytosis mediated by the TP63-RAC2 pathway promotes immunosuppressive remodeling in esophageal cancer
doi: 10.1016/j.xcrm.2025.102529
Figure Lengend Snippet: Efferocytosis and polarization of macrophages were analyzed in vitro (A) Western blotting verifies the differences in TP63 and RAC2 protein expression levels in the shTP63 and shTP63+RAC2 groups compared with the control group. (B) Differences in the fluorescence signal intensity of CMFDA green among groups of human primary macrophages with EC9706. ∗ p < 0.05. (C) Differences in the fluorescence signal intensity of CMFDA green among groups in THP-1-induced macrophages with EC9706. ∗∗ p < 0.01. (D) The fluorescence signal intensity of CMFDA green in human primary macrophages treated with EC9706 was determined by flow cytometry. ∗∗∗ p < 0.001. (E) The fluorescence signal intensity of CMFDA green in THP-1-induced macrophages treated with EC9706 was determined by flow cytometry. ∗∗∗ p < 0.001. (F) The CD163 + cell level of each group in human primary macrophages with EC9706. ∗∗ p < 0.01, ∗∗∗ p < 0.001. (G) The CD163 + cell level of each group in THP-1-induced macrophages with EC9706. ∗∗∗ p < 0.001. (H) The CD86 + cells in each group of human primary macrophages treated with EC9706. ∗∗ p < 0.01, ∗∗∗ p < 0.001. (I) The CD86 + cells in each group of THP-1-induced macrophages treated with EC9706. ∗∗∗ p < 0.001. (J) Differences in IL-10, TGF-β, and TNF-α levels among the groups in human primary macrophages with EC9706. ∗∗∗ p < 0.001. (K) Differences in IL-10, TGF-β, and TNF-α levels among the groups in THP-1-induced macrophages with EC9706. Data are presented as the mean ± SD of five independent experiments. ∗∗∗ p < 0.001.
Article Snippet: Then, slides were treated by microwave to induce antigen retrieval using citric acid solution for 15 min. For mIHC analysis of human samples, a panel of primary antibodies were used, including CD68 (1:100, ab283654, Abcam), Annexin A1 (1:1000, ab214486, Abcam), TP63(1:200, ab124762, Abcam), RAC2(1:2000,60077-1-Ig, Proteintech).
Techniques: In Vitro, Western Blot, Expressing, Control, Fluorescence, Flow Cytometry