Fig. 6: Obesity and Wnt5a expression in fat specific Wnt5a knock-out mice.

A Western blot analyses of Wnt5a protein in mouse serum (left panel). Ponceau S staining was performed to confirm equal loadings. Serum was obtained from the hearts of three male C57BL/6 J mice fed the ND or the HFD for 20 weeks from 8 weeks old. Relative band intensities of Wnt5a were obtained using ImageJ software and normalized versus Ponceau S band intensities (right panel). B–D Fat specific Wnt5a knock out (FKO) mice were generated by disrupting Wnt5a exon 2, as described in Supplementary Fig. S1A. B qRT-PCR of Wnt5a mRNA levels normalized versus 18 S rRNA levels in liver, subcutaneous WATs, or epididymal WATs of male Wnt5a^fl/fl mice (Floxed), and male FKO mice fed the ND for 20 weeks from 8 weeks old (Floxed 20 w ND, n = 5; FKO 20 w ND, n = 4). C Western blot analyses of Wnt5a protein in the sWATs of Floxed and FKO mice fed the ND or the HFD for 20 weeks from 8 weeks old. 14-3-3γ was used as the loading control. D Relative band intensities of Wnt5a were determined using ImageJ software and normalized versus 14-3-3γ band intensities. E Concentration of Wnt5a protein in the sera of Floxed and FKO mice as determined using a Mouse Wnt5a ELISA kit (AVIVA). Floxed and FKO mice were fed the ND or the HFD for 20 weeks from 8 weeks old (Floxed ND, n = 5; Floxed HFD, n = 6; FKO ND, n = 4; FKO HFD, n = 6). The significances of differences were assessed using a two-tailed paired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant.