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: International Dental Journal

Article Title: Immunological Features of Peri-Implant Soft Tissue After Healing Abutment Dislodgement: A Comparative Human Study

doi: 10.1016/j.identj.2026.109521

Figure Lengend Snippet: Immunohistochemical staining for immune cell markers in the epithelial + superficial connective tissue region. Representative immunohistochemical images of the superficial peri-implant soft tissue zone stained for MPO, CD68, CD3, CD20, and CD138. Scale bars: 100 μm. Red dashed boxes and red arrows indicate the magnified regions (insets).

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies (MPO, CD68, CD3, CD20 and CD138, all purchased from Boster, China; dilution 1:400).

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Molecular Medicine

Article Title: ZEB1 maintains mitochondrial fission and macrophage efferocytosis by restraining MFN2, thereby limiting inflammation and improving tendon-bone healing

doi: 10.3892/ijmm.2026.5805

Figure Lengend Snippet: ZEB1 knockdown aggravates the inflammatory response at the TBI. (A) The expression level of ZEB1 was detected by RT-qPCR 48 h after the operation. (B) The expression of ZEB1 protein was detected by IHC 1 week after the operation, and the optical density value was semi-quantitatively analyzed by ImageJ software (n=3 high-power fields) (scale bar, 50 µ m; lower panels, ×50 magnification, scale bar, 20 µ m). (C) Apoptotic cells at the TBI were detected by TUNEL method in the early postoperative period (1 week), with green fluorescence (FITC labeled) indicating apoptotic cells with DNA fragmentation (scale bar, 20 µ m). (D) The expression level of MFN2 was detected by RT-qPCR 48 h after the operation. (E) The expression of MFN2 protein was detected by IHC 1 week after the operation, and the optical density value was semi-quantitatively analyzed by ImageJ software (scale bar, 50 µ m). (F) At 1 week after the operation, double immunofluorescence staining was used to detect the polarization phenotype of macrophages. The cell nuclei were stained with DAPI (red), M1 macrophages were labeled with iNOS (green), and M2 macrophages were labeled with CD206 (red) (scale bar, 20 µ m) (G) At 1 week after the operation, co-localization analysis of apoptotic cells (Tunel, green) and macrophages (CD68, red) at the TBI was performed by immunofluorescence. The cell nuclei were stained with DAPI (blue) (scale bar, 20 µ m). The labels in the figure are as follows: T=tendon, B=bone tissue, I=tendon-bone interface. n=3. * P<0.05 vs. NC group. TBI, tendon-bone interface; ZEB1, zinc finger E-box binding homeobox 1; RT-qPCR, reverse transcription-quantitative PCR; IHC, immunohistochemistry; MFN2, Mitofusin-2; iNOS, inducible nitric oxide synthase; NC, negative control; sh, short hairpin.

Article Snippet: Then, the sections were incubated overnight at 4°C with primary antibodies against iNOS (1:50; cat. no. 18985-1-AP), CD206 (1:50; cat. no. 18704-1-AP) and CD68 (1;50; cat. no. 28058-1-AP) (Proteintech Group, Inc.) according to the manufacturer's instructions.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Software, TUNEL Assay, Fluorescence, Labeling, Double Immunofluorescence Staining, Staining, Immunofluorescence, Binding Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Immunohistochemistry, Negative Control

Journal: Journal of Advanced Research

Article Title: CCR1 hi /CCL5 hi macrophage-mediated CCL5 hi T cell chemotaxis in salivary gland aggravates Sjögren’s syndrome

doi: 10.1016/j.jare.2025.06.076

Figure Lengend Snippet: CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 and CCL5 expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) ELISA measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.

Article Snippet: To induce differentiation into macrophages, THP-1 cells were treated with 100 nM PMA for 24 h. Differentiation was confirmed by immunofluorescence analysis of CD68 localization and expression using a mouse anti-CD68 antibody (1:500, 66231–2-Ig, Proteintech, Wuhan, China).

Techniques: Migration, Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Transfection, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay