Journal: European Journal of Medical Research

Article Title: NDC80 promotes epithelial to mesenchymal transition of esophageal squamous cell carcinoma through macrophages polarization and PI3K/AKT pathway activation

doi: 10.1186/s40001-025-03397-3

Figure Lengend Snippet: NDC80 modulated tumor-associated macrophages polarization to promote ESCC progression. ( A ) Immunohistochemistry was used to detect the expression of NDC80 protein and macrophage markers in ESCC tumor tissue, n = 61. B, C Correlation between the expression of NDC80 protein and the number of macrophages infiltrations in ESCC tumor tissue was analyzed by simple linear regression, n = 61. Flow cytometry ( D ) and qPCR ( E ) were used to detect the regulation of macrophages polarization by ESCC cells overexpressing NDC80, n = 3, multiple unpaired t tests. F qPCR was used to detect the regulation of macrophages polarization by ESCC cells with silenced NDC80, n = 3, multiple unpaired t tests. G, H qPCR was used to assay the mRNA expression levels of inflammatory cytokines in tumor cells with different NDC80 expression levels, n = 3, multiple unpaired t tests. I, J ELISA detection of M-CSF and CXCL-2 levels in the supernatant of ESCC cells with overexpression or knockdown of NDC80 protein, n = 3, multiple unpaired t tests. M-CSF: Standard curve range = 31.25—2000 pg/mL, R 2 = 0.9997. CXCL-2: Standard curve range = 15.63—1000 pg/mL, R 2 = 0.9986. (*, p < 0.05; **, p < 0.01; ***, p < 0.001)

Article Snippet: The concentrations of cytokines M-CSF (E-EL-H0097) and CXCL-2 (E-EL-H1904) in the cell supernatants were detected using ELISA kits (Elabscience, China).

Techniques: Immunohistochemistry, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Over Expression, Knockdown

Journal: Advanced Science

Article Title: FAPα + Macrophages Orchestrate Immune Evasion in Multiple Myeloma by Dual Regulation of PD‐L1 and T Cell Senescence

doi: 10.1002/advs.202506239

Figure Lengend Snippet: FAPα + Macrophages Are Enriched in the Myeloma Microenvironment and Correlate with Disease Progression. (A) Heatmap of gene expression in macrophages from patients with MM (n = 3) or healthy donors (HDs, n = 3). (B,C) Cell percentages of FAPα + macrophages (CD11b + CD14 + ) and FAPα + BMSC (CD45 − CD38 − CD29 + ) in PBMCs or BMMCs from patients with MM (n = 9). (D) Reprehensive immunofluorescence staining (IF) images of CD138, CD68, and FAPα in the BM of a NDMM patient (Magnification ×400. Scale bar, 50 µm). (E–G) Flow cytometry analysis of the cell percentages of FAPα + macrophages and FAPα + BMSCs in BMMCs from patients with MM at different disease stages (n = 27) or HDs (n = 4). (H) Correlation analysis of the cell percentages between CD138 + MM cells and FAPα + macrophages or FAPα + BMSCs in patients with NDMM (n = 15). (I) Reprehensive immunohistochemical staining (IHC) images of CD138 and FAPα in patients with NDMM or MM‐CR (Magnification ×200. Scale bar, 50 µm). (J,K) Western blot (J) and flow cytometry (K) analysis of FAPα expression in macrophages co‐cultured with MM cells (n = 3). (L) TGFβ1 expression in different MM cell lines. (M‐N) TGFβ1‐induced FAPα protein expression in macrophages (Magnification ×400. Scale bar, 50 µm). (O) M‐CSF, TGFβ1, and FAPα levels in BM supernatants from patients with MM at different disease stages (n = 37) using ELISA assay. (P) Correlation analysis of FAPα and TGFβ1 or M‐CSF in BM supernatant from patients with NDMM (n = 17). (Q) Reprehensive IHC images of bone marrow samples from patients with MM (n = 18) (Magnification ×200. Scale bar, 50 µm). (R) IOD values of CD138, CD68, and FAPα in patients with NDMM or RRMM (n = 18). (S) Correlation analysis between CD138 and FAPα in patients with NDMM. (T) Kaplan–Meier curves of PFS and overall OS in the set of patients with NDMM based on FAP protein expression level detected in tumor tissues. The median value of FAP RNA expression in the was 88.26 (IOD). The expression value of the FAP high group (n = 9) was >88.26(IOD) and the FAP low group (n = 9) was <88.26(IOD). Data are presented as mean ± SD. Each dot means independent samples. ns, no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Statistical analysis was performed using a 2‐tailed Student's t ‐test in C, E, F, G, K, O, and R, a Pearson correlation in H, P, and S, a log‐rank test in T. MM, multiple myeloma; HD, healthy donors; BMSCs, bone marrow mesenchymal stem cells; PBMCs, peripheral blood mononuclear cells; BMMCs, bone marrow mononuclear cells; BM, bone marrow; NDMM, newly diagnosed MM; RRMM, relapsed or refractory MM; CR, complete response; TGFβ1, Transforming growth factor beta 1; M‐CSF, macrophage colony stimulating factor; IHC, Immunohistochemistry; IOD, Integrated Optical Density; RRMM, Relapsed/Refractory MM; PFS, Progression‐free survival; OS, overall survival.

Article Snippet: The bone marrow supernatants of patients with different MM stages were collected, and different cytokines were detected as described by the respective manufacturers: Human M‐CSF AccuSignal ELISA Kit (Cat #KOA0253, Rockland, USA), Human Seprase/FAP AccuSignal ELISA Kit (Cat #KOA0627, Rockland, USA), Human TGFβ1 ELISA Kit (Cat #1117102) (All from Dakewe Bioengineering Co., Ltd, China).

Techniques: Biomarker Discovery, Gene Expression, Immunofluorescence, Staining, Flow Cytometry, Immunohistochemical staining, Western Blot, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, RNA Expression, Immunohistochemistry

Journal: BMC Musculoskeletal Disorders

Article Title: The link between osteoporosis and frozen shoulder: exploring the therapeutic effect of TAK715 on reversing fibrosis and protecting against osteoporosis via the p38 MAPK signaling pathway

doi: 10.1186/s12891-024-08068-8

Figure Lengend Snippet: GSEA and immune infiltration analysis after TAK715 treatment. A The selected KEGG GSEA analysis ES diagram: apoptosis (pathway ID: hsa04210), TNF (pathway ID: hsa04668), IL-17 (pathway ID: hsa04657), MAPK signaling pathway (pathway ID: hsa04010) and TGF-beta signaling pathway (pathway ID: hsa04010). The combined ES diagram was at last. B The combined heatmap of selected pathways in KEGG GSEA analysis. C The selected GO GSEA analysis ES diagram: The collagen binding (GO:0005518), collagen-containing extracellular matrix (GO:0062023), collagen fiber organization (GO:0030199), growth factor activity (GO:0008083) and collagen trimer (GO:0005581). The combined ES diagram was at last. D The combined heatmap of selected pathways in KEGG GO analysis. E The inflammation-collagen interaction chord gram showed that the collagen terms and inflammation pathways are tightly linked. The value between term and pathway was calculated according to the log 2 fold change of common genes. F The heatmap of different immune-cell gene expression in this gene set. The M1 macrophage related genes were more expressed in untreated FS synovial fibroblasts (NC group) compared with TAK715 treated (TAK5 group). G The correlation and p value of immune infiltration result. The M1 macrophage cluster exhibited the correlation of 0.18 and p value of 0.32, which indicated that the M1 macrophage infiltration was positive linked with the FS synovial fibroblasts

Article Snippet: The bone marrow derived macrophages (BMDMs) were cultured according to previous studies and seeded in a six-well plate with 5 × 105 per well and stimulated with rat macrophage colony stimulating factor (M-CSF, MCE, NJ, USA) at 30 ng/ml.

Techniques: Binding Assay, Activity Assay, Gene Expression

Journal: BMC Musculoskeletal Disorders

Article Title: The link between osteoporosis and frozen shoulder: exploring the therapeutic effect of TAK715 on reversing fibrosis and protecting against osteoporosis via the p38 MAPK signaling pathway

doi: 10.1186/s12891-024-08068-8

Figure Lengend Snippet: the X-ray images ( A , B ), ROM measurement ( C , D ) and histology analysis of FS rat models. A , B The X-rays of FS rats in neutral and abduction position and the statistics analysis of angle measurement in ( A ). ( n = 6) * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. C , D The ROM measurement of FS rats in neutral and abduction position and the statistics analysis of ROM measurement in ( A ). ( n = 6) * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. E Representative hematoxylin and eosin-stained (H&E) tissue sections of the shoulder joint. The capsule regions were encircled in dotted rectangle. Scale Bar: 100 μM. F Representative masson-stained tissue sections of the shoulder joint. The capsule regions were encircled in dotted rectangle. Scale Bar: 100 μM. G The statistics analysis of capsular thickness(mm) and macrophage relative counts in ( E ). ( n = 3) * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The bone marrow derived macrophages (BMDMs) were cultured according to previous studies and seeded in a six-well plate with 5 × 105 per well and stimulated with rat macrophage colony stimulating factor (M-CSF, MCE, NJ, USA) at 30 ng/ml.

Techniques: Staining

Journal: BMC Musculoskeletal Disorders

Article Title: The link between osteoporosis and frozen shoulder: exploring the therapeutic effect of TAK715 on reversing fibrosis and protecting against osteoporosis via the p38 MAPK signaling pathway

doi: 10.1186/s12891-024-08068-8

Figure Lengend Snippet: The pathogenesis mechanism based on the activation of p38 MAPK signal pathway and the therapeutic effect of TAK715. Briefly, the extrinsic or intrinsic reasons produced fibrogenic stimuli and the stimuli triggered synovium fibroblast apoptosis and recruited macrophage infiltration. At the initial pathogenesis phase, the osteoclast was also activated. In this stage, the patients suffered from pain, stiffness and bone loss. Under the cessation of fibrogenic stimuli and consistent cell apoptosis, the ECM secreting is stopped and degraded. The FS comes into thawing phase. Holistically, the TAK715 could correct the fibrosis and local osteoporosis to accelerate the rehabilitation. (Created from https://www.biorender.com )

Article Snippet: The bone marrow derived macrophages (BMDMs) were cultured according to previous studies and seeded in a six-well plate with 5 × 105 per well and stimulated with rat macrophage colony stimulating factor (M-CSF, MCE, NJ, USA) at 30 ng/ml.

Techniques: Activation Assay, Produced

Journal: Journal of Advanced Research

Article Title: The fatty acid receptor CD36 promotes macrophage infiltration via p110γ signaling to stimulate metastasis

doi: 10.1016/j.jare.2024.10.006

Figure Lengend Snippet: CD36 mediates the recruitment of macrophages through CCL2/CCR2 axis. A, The mRNA levels of chemokines in normal liver or LLC metastatic liver measured by RT-PCR (n = 4). c p < 0.05, b p < 0.01, a p < 0.001 vs normal liver. B, C, The mRNA levels of chemokines (B) and chemokine receptors (C) in tumor cells, BMDMs, CD11b + myeloid cells and blood monocytes (n = 4–6). c p < 0.05, b p < 0.01, a p < 0.001 vs the LLC group. D, E, The mRNA levels of Ccl2 or Mcsf (D) and their receptor Ccr2 or Csf1r (E) were measured in WT and Cd36 MKO mice with liver metastasis by RT-PCR (n = 5–6). F, G, The Ccr2 or Csf1r gene expression in WT and Cd36 -/- BMDMs (F), or in NC and CD36 OE THP-1 cells (G) cultured with TCM (n = 4–6). H, I, The MFI of CCR2 was measured in macrophages (H) and monocytes (I) isolated from metastatic liver tumors of WT and Cd36 MKO mice (n = 5). J, Migration assay was performed in WT or Cd36 -/- BMDMs treated with or without CCL2 or MCSF (n = 5). K-O, WT (n = 5) and Cd36 MKO (n = 5) mice were injected with LLC cells, following intraperitoneal injection of CCR2 inhibitor, RS504393, once a day from the 4th day to 14th day. HE-stained liver tissues along with the quantification of tumor area from WT and Cd36 MKO mice after treated with RS504393 (K, M, n = 3). F4/80-stained metastatic liver tissues from WT and Cd36 MKO mice after treated with RS504393 (L, M, n = 5). TSNE analysis of CD45 + cells isolated from the liver based on FCM (N). The percentage of indicated cells was measured (O, n = 5). Values for n represent biologically independent samples. Values for n represent biologically independent samples. Data are presented as mean ± SEM. *p < 0.05, ** p < 0.01, and *** p < 0.001 vs the control group; # p < 0.01, ## p < 0.01, and ### p < 0.001 vs the WT group. Unpaired two-tailed t -test (A, D-J), Unpaired two-tailed t -test with Mean-Whitney test (O) and ANOVA (B, C, M).

Article Snippet: Murine bone marrow-derived macrophages (BMDMs) were flushed from the tibia and femur of 6 ∼ 8-week-old WT or Cd36 MKO mice and differentiated in RPMI1640 containing M−CSF (10 ng/ml, #HY-P700137AF, MedChemExpress) for 5–7 days.

Techniques: Reverse Transcription Polymerase Chain Reaction, Gene Expression, Cell Culture, Isolation, Migration, Injection, Staining, Control, Two Tailed Test

Journal: iScience

Article Title: Identification of potential biomarkers and therapeutic targets for antineutrophil cytoplasmic antibody-associated glomerulonephritis

doi: 10.1016/j.isci.2023.108157

Figure Lengend Snippet:

Article Snippet: Human M-CSF ELISA Kit , Boster , EK0444.

Techniques: Enzyme-linked Immunosorbent Assay, Software