Soluble extracts were prepared by resuspending whole cells in the corresponding extraction buffer [32], cells were lysed with glass beads.
Cells in the lysis buffer described above were lysed with three repeating cycles of alternating one-min incubations from the ethanol/dry ice mix to a 37°C water bath before RT-PCR.
After washing with phosphate-buffered saline, they were lysed with RLT buffer (Qiagen, Hilden, Germany) and frozen at −70°C.
After washing with phosphate-buffered saline, cells were lysed with RLT buffer (Qiagen, Hilden, Germany) and frozen at -70°C.
After 7 days, cells were lysed with buffer containing 0.5%% (v/v) Triton X-100, and adipose conversion was quantitated by the activity of the adipogenic enzyme glycerophosphate dehydrogenase.
Cells were lysed with buffer containing 1% Triton X-100, 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM CaCl2, and a protease inhibitor cocktail (Roche).
After 24 h of transfection, HAoSMCs were lysed with Buffer RLT (Qiagen, Limburg, Netherlands).
Cells were lysed with buffer containing 100 mM Tris-HCl (pH 7.4) and 4% sodium dodecyl sulfate.
For Western blot, cells were lysed with SDS buffer or NP40 in NET buffer.
Then those cells were lysed with buffers and incubated with 40 µg of GST-RBD (Cytoskeleton, Acoma Street, Denver, USA) for 1 hour.
