Human fgf1

Mice were housed in a temperature-controlled environment with a 12-hr light/12-hr dark cycle and handled according to Salk IACUC guidelines complying with U.S. legislation. Male ob/ob mice (B6.V-Lep^ob/J, Jackson laboratories) and male C57BL/6J mice received a standard or high fat diet (MI laboratory rodent diet 5001, Harlan Teklad; high fat (60%) diet F3282, Bio-Serv) and acidified water ad libitum. STZ-induced diabetic mice in the C57BL/6J background were purchased from Jackson laboratories. aP2-Cre mice (B6.Cg-Tg(Fabp4-cre)1Rev/J, Jackson laboratories) were crossed to FGFR1 floxed mice (B6.129S4-Fgfr1tm5.1Sor/J, Jackson laboratories) to generate aP2-Cre; FGFR1 fl/fl mice. 0.1 mg/ml solutions in PBS of mouse FGF1 (Prospec, Ness Ziona, Israel), human FGF1 (Prospec, Ness Ziona, Israel), mouse FGF2 (Prospec, Ness Ziona, Israel), mouse FGF9 (Prospec, Ness Ziona, Israel), and mouse FGF10 (R&D systems) were injected as described. Heparin sodium salt (Sigma) was premixed with mFGF1 peptide prior to injection.

A heparin-Sepharose column (100 μL, GE Healthcare) was pre-equilibrated with PBS (Gibco) then loaded with human FGF1 (ProSpec), FGF2 (Peprotech), FGF7 (ProSpec), or FGF10 (ProSpec) dissolved in PBS. The flow-through was reloaded onto the column twice to ensure complete binding. After washing twice with PBS, a 0.6 mg/mL solution of Sulfo-NHS-LC-biotin (Thermo Fisher) in PBS was loaded onto the column and incubated for 1 hr at room temperature. Each column was washed three times with PBS, then bound biotinylated protein was eluted with 0.4 mL of PBS buffer containing an additional 2 M NaCl. Biotinylated S100A12 was prepared as previously reported^{57 (link)}. All biotinylated proteins were stored at −80°C before use.