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: 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