Faeces in mixture buffer were disrupted with 0.1 mm zirconia/silica beads (BioSpec Products, Inc., USA) by vigorous shaking (1500 rpm, for 10 min) using a Shake Master Biomedical Sciencee).
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).
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).
After adding the fragmentation buffer, mRNA was disrupted into short fragments (≈200 bp).
