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: Cell Death & Disease

Article Title: Melatonin attenuates detrimental effects of diabetes on the niche of mouse spermatogonial stem cells by maintaining Leydig cells

doi: 10.1038/s41419-018-0956-4

Figure Lengend Snippet: a CSF1 expression with Hoechst 33342 and their merges of testicular sections from mice under four different treatments in the short term (W2, 2 weeks) and long term (W8, 8 weeks) experiments. Scale bar, 200 μm. b ELISA of CSF1 in mice serum of the four different groups in W2 and W8 experiments. c RT-qPCR detected Csf1 expression in mice testicular tissue under the four different treatments in W2 and W8 experiments. d CSF1 expression level detected by western blot analysis in the four groups in W2 and W8. Histograms are the statistical result of Image J (V1.48d) gradation analysis for the western blot experiments. (The results are shown as the mean ± S.E.M of at least three mice ( n = 3) and the statistical significance was expressed as follows: * p < 0.05; ** p < 0.01). C control group, M normal group treated with MLT, D diabetic group, DM MLT-treated diabetic group

Article Snippet: Mouse CSF1 level of mouse plasma was quantified using an ELISA kit (Boster, Wuhan, China) according to the instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Melatonin attenuates detrimental effects of diabetes on the niche of mouse spermatogonial stem cells by maintaining Leydig cells

doi: 10.1038/s41419-018-0956-4

Figure Lengend Snippet: a CSF1 expression with Hoechst 33342 and their merges of MLTC-1 under three different ERS states (C control, Tm Tm treatment induced excessive ERS, Tm + 4PBA Tm treatment in combination with 4PBA for alleviated ERS). b CSF1 concentration in culture medium of MLTC-1 under the three different treatments was detected using ELISA. c RT-qPCR detected the mRNA expression level of Csf1 in MLTC-1 under the three treatments. d Expression of CSF1 in MLTC-1 under different treatments was measured by Western blot. Histograms are the statistical result of Image J (V1.48d) gradation analysis for the western blot experiments. e A graphical conclusion of this study is displayed: DM-induced high glucose activated ERS response factors (Grp78 and CHOP) in Leydig cells, resulting in apoptosis of Leydig cells and growth arrest of SSCs in testes. In response to these changes, melatonin treatment alleviated the apoptosis of Leydig cells via inhibition of ERS and recovered the CSF1 secretion in testes. CSF1 then resumed the self-renewal capacity of SSCs. (The results are expressed as the mean ± S.E.M of three separated wells of cells ( n = 3) in at least three different experiments and the statistical significance is expressed as follows: * p < 0.05; ** p < 0.01)

Article Snippet: Mouse CSF1 level of mouse plasma was quantified using an ELISA kit (Boster, Wuhan, China) according to the instructions.

Techniques: Expressing, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Inhibition

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