Sis3 hcl

Each rat (n = 9 in each group) received SIS3-Hcl (Selleck, SIS3 in short) disolved in 10% DMSO by intraperitoneal injection (2.8 mg/Kg) 7 days before syringomyelia induction. 10% DMSO was injected as control. DMSO and SIS3 injections were continued until the end of the experiment. All of the rats in the two groups were not decompressed after syringomyelia induction.

First, 3T3-L1 cells were differentiated into adipocytes using differentiation medium containing 3-isobutyl-1-methylxanthine (0.5 mmol/l), dexamethasone (1 μmol/l), insulin (1 μmol/l), and pioglitazone (10 μmol/l). The cells were used in experiments 7 days after differentiation. Treatment with recombinant human TGF-β1 (R&D Systems) was performed after changing the media to serum-free Dulbecco's modified Eagle's medium to avoid unknown serum effects. PlatE cells were used to produce retrovirus, followed by transfection in 3T3-L1 cells. Stable 3T3-L1 cells expressing tetracycline (tet)-inducible TGF-β1 (3T3-L1–tet–TGF-β1) or control cells (3T3-L1–tet–empty cell) were produced using the Retro-X Tet-On Advanced system according to the protocol supplied by the manufacturer (Clontech). The coding region of mouse TGF-β1 was subcloned into the expression vector pRetroX-Tight-Pur. Retroviral particles were generated using pRetroX-Tight-Pur-TGF-β1 or pRetroX-Tight-hygro-empty and pRetroX-tet-on advanced vectors. Infected 3T3-L1 cells were selected in 400 mg/ml G418 and 5 mg/ml puromycin. Other materials were as described: SB431542 (Selleck Chemicals), SIS3 HCl (Selleck), PF-573228 (Selleck), MK2206 (Selleck), collagenase (Sigma–Aldrich), rapamycin (Selleck), and U0126 (Selleck).