Journal: Current Issues in Molecular Biology

Article Title: Coptis chinensis Franch. Suppresses Invasive Pulmonary Aspergillosis by Augmenting NADPH-Dependent Neutrophil Extracellular Traps via Dual Modulation of Complement Activation and Gut Microbiota

doi: 10.3390/cimb48040424

Figure Lengend Snippet: Bayesian networks and KEGG suggest a CCF-biased reorganization toward CR3-SYK-NOX/NET pathways relative to IPA/AmB. The network analysis highlights recurrent CCF-associated hub genes, including Itgam, Syk, Rac2, and Cybb, together with pathway enrichment linked to oxidative and NET-related responses. ( A ) Hub gene networks identified in the IPA, CCF, and AmB groups by Bayesian network analysis. Genes were ranked by DEGREE value, with the top 10% defined as hub genes and marked in color; nodes with DEGREE >6 were placed at the core of the network. ( B ) KEGG pathway enrichment of hub genes in each group. Pathways were ranked by −log 10 ( p value). Bar colors represent different groups: IPA (dark blue), CCF (dark red), and AmB (light blue). The red box denotes the most significantly enriched KEGG pathway in the CCF group. ( C ) FPKM values of key CCF-regulated genes at +2, +4, and +6 dpi. Statistical analysis was conducted comparing all groups to the CCF group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 indicate levels of statistical significance. ( D ) qPCR validation of key CCF-regulated genes in each group. Statistical analysis was conducted using one-way ANOVA, comparing all groups to the CCF group. * p < 0.05, ** p < 0.01, **** p < 0.0001 indicate levels of statistical significance. Data are presented as mean ± SD from n = 3 mice per group.

Article Snippet: Primary antibodies used in Western Blot included Beta Actin Monoclonal antibody (66009-1-Ig, Proteintech, Wuhan, China), NCF4/p40phox Rabbit pAb (A2096, Abclonal, Wuhan, China), Anti-NOX2/gp91phox Recombinant Rabbit Monoclonal Antibody (ET1611-44, Huabio, Hangzhou, China), RAC2 Rabbit pAb (A1139, Abclonal, China) and Anti-NCF1/p47phox Rabbit pAb (GB11724, Servicebio, Wuhan, China).

Techniques: Biomarker Discovery

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

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: Experimental study of shTP63 and shTP63+RAC2 in apoptotic AKR (apoptotic esophageal carcinoma) cells-C57BL/6 efferocytosis mouse model (A) Western blot verifies the difference in TP63 and RAC2 protein expression levels in shTP63 and shTP63+RAC2 groups compared with the control group. (B) The efferocytosis index level of each group was detected ( n = 5 per group). ∗∗ p < 0.01, ∗∗∗ p < 0.001. (C) The CD163 + cells in each group were detected ( n = 5 per group). ∗ p < 0.05, ∗∗ p < 0.01. (D) The CD86 + cells in each group ( n = 5 per group). ∗∗ p < 0.01, ∗∗∗ 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: Western Blot, Expressing, Control