Half of the cell suspension in PBS buffer was disrupted with the ultrasonic homogenizer, while the remaining suspension was left on ice as intact cells.
After washing twice and resuspending with 50 mM phosphate buffer (pH 7.0), the cells were disrupted with an ultrasonic cell breaking apparatus (Xinzhi, Ningbo, China).
The cell membranes were disrupted with Dounce buffer, which causes swelling of the cells, and a fine tissue homogenizer that enables release of the nucleus without destroying it.
VSMCs (5 × 10 cells/cm) were disrupted with lysis buffer (50 mmol/l Tris-HCl (pH 8.0), 150 mmol/l NaCl, 0.02% sodium azide, 100 µg/ml phenylmethylsulfonyl fluoride, 1 µg/ml aprotinin, 1% Triton X-100).
For Western blotting on intracellular Salmonella cells, infected macrophage cells were disrupted with a lysis buffer (50 mM HEPES, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 100 mM PMSF, protease inhibitor cocktail (Roche), 50 mM NaF, DNase I (Qiagen; DNase Set), and 2 mM sodium orthovanadate) on ice for 10 min and centrifuged at 10,000 × g for 10 min to pellet intracellular bacteria.
After DNP treatment, the cell samples were disrupted with 0.6 mL lysis buffer.
Cell pastes of 6H2N-induced C. guilliermondii and recombinant E. coli were disrupted with glass beads in the extraction buffer.
The resulting cell pellets were disrupted with 100-μm glass beads (Sigma) in pre-chilled buffer, appropriate for the subsequent assays, using a Precellys®24 bead beater (Bertin Technologies).
Two days after transient transfection, 293T cells were harvested and washed with ice-cold phosphate-buffered saline before being disrupted with lysis buffer (50 mM Tris, 100 mM NaCl, 0.1% NP-40, 50 mM NaF, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride, 1 μg/ml of aprotinin, 0.5 μg/ml of leupeptin, and 0.7 μg/ml of pepstatin).
To extract the genomic DNA of the isolated microorganisms, cells were suspended in 100 μl each of Tris-EDTA (TE -saturated phenol and TE -saturatedd were disruphenoly beandng wiTE zirconia buffer(Wen et and 2005).
