Fig 1: Slit2 is an osteokine secreted by osteoblasts that protects mice from DIO.a Heatmap showing relative RNA levels of several cytokines in primary osteoblasts isolated from WT and Shn3−/− mice (n = 3). b Immunoblotting for SLIT2 in osteoblast lysates, serum, osteoblast supernatant. The red and bold blue boxes indicate regions that were cut for further LC-MS studies. Results were selected from 3 repeated experiments. c Matched peptides of SLIT2 and the related amino acid sequence detected in murine sera. d ELISA measurement of SLIT2 in conditioned media from WT and Shn3−/− osteoblasts (n = 4 for each group). (e) Immunoblotting demonstrating the levels of SLIT2-C in the serum of HFD-fed Shn3−/− and WT mice and (f) ELISA measurement of circulating SLIT2 levels in Shn3−/− and WT mice after 12-weeks of HFD exposure (n = 9 for WT group and n = 11 for Shn3−/−group). g Immunoblotting for SLIT2-C in the serum of HFD-exposed Slit2Osx and Slit2f/f mice. h ELISA results of circulating SLIT2 levels in Slit2f/f and Slit2Osx mice (n = 6 for each group). i Body weight curve of HFD-exposed Slit2Osx and Slit2f/f mice (n = 6 for each group). j Representative gross images of iWAT obtained from Slit2Osx and Slit2f/f mice. k tissue wet weight of iWAT, eWAT, liver and BAT obtained from Slit2Osx and Slit2f/f mice (n = 6 for each group). l Representative histological sections of iWAT, eWAT and liver from 3 individual slides of Slit2Osx and Slit2f/f mice after a 10-week-HFD challenge. Scale bar=100 um. m Thermogenic gene expression level relative to Gapdh in iWAT of HFD-fed Slit2Osx and Slit2f/f mice (n = 4 for each group). Results were presented as mean ± s.d., analyzed by unpaired, two-tailed Student’s t test (e, f, h, k and m) or two-way ANOVA with Bonferroni’s multiple comparisons test (i). *P < 0.05. Source data are provided as a Source Data file.