Figure 1
Sema3E vaccine inhibits inflammation in visceral fat. (A) Mice fed a high fat diet (HFD) were immunized with KLH-conjugated HKEGPEYHWS peptide (Sema3E vaccine group) or KLH alone (KLH group), and plasma antibody titers were analyzed with ELISA at 6, 10, 14 and 20 weeks of age (n = 4, 4, 4, 4, 4, 4, 4, 4). (B) After the third dose, plasma was collected from the Sema3E vaccine group and the KLH group (Con). Recombinant Sema3E (r-Sema3E) protein or BSA-conjugated Sema3E peptide (Pep) was blotted with plasma samples or with a commercially available anti-Sema3E antibody (Sema3E-Ab). (C) Hematoxylin and eosin (HE) staining of epididymal WAT (eWAT) harvested from mice in the Sema3E vaccine group (Sema3E vaccine) or the KLH group (Con) maintained on a HFD. Nos. 1, 2, and 3 indicate that the eWAT originated from different mice. Scale bar = 50 μm. (D–F) Levels of transcripts for Adgre1 (Emr1)(D) (n = 6, 6), Plxnd1 (E) (n = 6, 6), Itgam (n = 6, 6), and Itgax (n = 6, 6)(F) in the indicated groups. (G) Evaluation of reactive oxygen species (ROS) performed by dihydroethidium (DHE) staining in the indicated groups. Scale bar = 50 μm. (H) Western blot analysis of p53 in the indicated groups. Full-length blots are presented in Supplementary Fig. 2F. (I) Cdkn1a expression was examined in the indicated groups (n = 6, 6). Outliers were excluded by boxplot analysis. All remaining data were analyzed by the two-tailed Student’s t-test. *P < 0.05, **P < 0.01. Values represent the mean ± s.e.m. NS = not significant.
