The frozen brain tissue samples were weighed and homogenized in acidified butanol.
The highest levels of S1PR2 in human brain tissue samples were in two female MS patients who had "relapsing and remitting MS", meaning their symptoms flared irregularly.
Harvested brain tissue samples were digested into single cell suspension using papain as described in the methods.
Frozen brain tissue samples were homogenized on ice using ultrasound in a 3-fold (w/v) volume of 1% (w/v) sulfosalicylic acid.
Briefly, brain tissue samples were homogenized in 2∶1 chloroform:methanol containing 0.5% butylated hydroxytolulene and lipids were extracted by the method of Folch [58].
The brain tissue samples were obtained from the 307 Hospital of PLA with the written informed consent of patients and with approval for experiments from the ethics committees of the hospital and the Beijing Institute of Biotechnology.
Brain tissue samples were lysed in radioimmunoprecipitation assay (RIPA) buffer [10 mmol/L Tris, 140 mmol/L NaCl, 1% Triton X-100, 1% Na deoxycholate, 0.1% SDS and protease inhibitor cocktail (Pierce #78410) pH 7.5] and centrifugated for 15 min at 10,000 RPM.
Brain tissue samples were analyzed by ELISA for SHH.
Brain tissue samples were prepared as described above.
Brain tissue samples were stored at −80°C until analysis.
Brain tissue samples were frozen and stored at −80 °C.
