Figure 4

Adipocyte-derived molecules that support eosinophil migration and survival account for eosinophil infiltration in the perigonadal adipose tissue of high fat diet (HFD) fed mice. (a) mRNA expression of CC chemokine ligand 11 (Ccl11) and Ccl24 in adipocyte-differentiated 3T3-L1 cells. Representative blots from 3 independent experiments. The full-length blots are presented in Fig. S6. ***p < 0.001 (Student’s t-test) (b) Production of CCL11 and CCL24 by cultures of adipocyte-differentiated 3T3-L1 cells. ***p < 0.001 (Student’s t-test) (c) Migration of eosinophils isolated from the peritoneal cavity of CD3-IL-5 transgenic mice toward adipocyte-differentiated 3T3-L1 cells treated with or without anti-mouse CCL11. ***p < 0.001 (Student’s t-test) (d) mRNA expression of interleukin (IL)-3 (Il3), IL-5 (Il5), and granulocyte-macrophage colony-stimulating factor (Csf2) in adipocyte-differentiated 3T3-L1 cells. Representative blots from 3 independent experiments. The full-length blots are presented in Fig. S6. (e) Viability of eosinophils isolated from the peritoneal cavity of CD3-IL-5 transgenic mice and co-cultured with or without adipocyte-differentiated 3T3-L1 cells. Non-viable eosinophils were detected by staining with 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI). ***p < 0.001 (Student’s t-test) (f) mRNA expression of Ccr3 in the perigonadal fat, liver, muscle, and small intestine (SI) of wild-type (WT) mice fed a HFD or chow. **p < 0.01 (Student’s t-test for fat, muscle, and SI, Mann-Whitney test for liver). (g) mRNA expression of Ccl11 in the perigonadal fat of WT mice fed a HFD or chow. ***p < 0.001 (Student’s t-test) (h) Flow cytometric analysis of CCR3⁺SiglecF⁺ eosinophils in the perigonadal fat and bone marrow (BM) of WT mice fed a HFD or chow. Representative dot plots are shown. **p < 0.01 (Student’s t-test). Graphs show the mean ± standard error of the mean.