Journal: Frontiers in Immunology

Article Title: Macrophage morphology in the tumor microenvironment predicts metachronous liver metastasis in gastric cancer: establishment and validation of a predictive model

doi: 10.3389/fimmu.2026.1770436

Figure Lengend Snippet: The study design is presented in six sequential steps. (1) Patient enrollment: a total of 1228 patients with gastric cancer who underwent curative gastrectomy between 2016 and 2020 were screened according to predefined inclusion and exclusion criteria. (2) PSM-based cohort construction: to reduce baseline imbalance between MLM and non-MLM groups, 1:2 propensity score matching (PSM) was performed, resulting in a matched cohort of 233 patients (86 MLM and 147 non-MLM cases). (3) Tissue staining and digitalization: formalin-fixed paraffin-embedded sections were stained with CD68 and CD163 antibodies, followed by whole-slide scanning. (4) QuPath-based digital pathology: tissue segmentation and annotation were performed in QuPath, and artificial neural network (ANN) learning was applied to extract macrophage morphological features (area and perimeter) in different histological regions. (5) Model development: the matched cohort was randomly divided into a 70% training set and a 30% validation set, and a predictive nomogram was constructed based on macrophage morphological parameters. (6) Model evaluation: the performance of the nomogram was assessed using receiver operating characteristic (ROC) analysis, calibration curves, and decision curve analysis (DCA). MLM, metachronous liver metastasis; IHC, immunohistochemistry.

Article Snippet: Subsequently, the slides were incubated overnight at 4 °C with primary antibodies targeting human CD68 (1:500, Servicebio, China) and CD163 (1:500, Servicebio, China).

Techniques: Staining, Formalin-fixed Paraffin-Embedded, Biomarker Discovery, Construct, Immunohistochemistry

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: MMP12 silencing inhibited M2 macrophage polarization. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. (A) Flow cytometry was used to quantify the number of CD68-positive cells. Subsequently, KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (B) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (C) Flow cytometry was used to quantify the number of CD206-positive macrophages. (D) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Quantitative RT-PCR, Migration, Transwell Migration Assay

Journal: Frontiers in Immunology

Article Title: The CXCL9/SPP1 polarity axis in tumor-associated macrophages: immunoregulatory and prognostic significance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1763652

Figure Lengend Snippet: Validation of CXCL9 + and SPP1 + macrophage expression and polarization characteristics in lung cancer tissues. (A) Representative mIF staining for CXCL9+ TAMs and SPP1+ TAMs in lung tissue sections from the normal and LUAD tumor groups. DAPI (blue), CD68 (red), CXCL9 (yellow), SPP1 (green) are shown, along with individual and merged channels. (n = 3 per group). Scale bar, 20 μm. (B) Representative mIF staining of CXCL9+ TAMs (yellow) and SPP1+ TAMs (green) in lung tissue sections from the normal and LUAD tumor groups (n = 3 per group). Scale bar, 20 μm. (C) qRT-PCR validation of macrophage polarization. The M1 group showed high expression of Cxcl9 and iNOS, while the M2 group showed high expression of Spp1 and Arg1. Asterisks indicate statistically significant differences (*p < 0.05).

Article Snippet: Following another stripping step, the third round was performed using mouse anti-human CD68 monoclonal antibody (1:50000, Servicebio, Cat# GB153150 ) with HRP-conjugated goat anti-mouse secondary antibody (ready-to-use, Servicebio, Cat# G1301), and signal was developed using iF555-Tyramide (1:500, Servicebio, Cat# G1233).

Techniques: Biomarker Discovery, Expressing, Staining, Quantitative RT-PCR