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