The harvested treated and non-treated cells after trypsinization were stained for flow cytometry at a concentration of 1×106 cells per 100 µl of buffer (PBS pH 7.4, 2% PBS, 2 mM EDTA).
From each of 6 sections in a field, oats were harvested, treated (INO) or not (CON) with inoculant, packed into 19-L BKT or vacuum bags (BG), and ensiled for 217 d.
LPS blasts were harvested, treated with mitomycin- C (50 µg/ml) for 20 mins at 37°C, and pulse-labeled with OVA257 264 peptide or NP 366 374 peptide at a final concentration of 10 µg/ml for 1 h at 37°C in a water bath.
After metaphase arrest through exposure to Colcemide, cells were harvested, treated with hypotonic solution, and then fixed with methanol and acetic acid according to standard procedures.
On the day of harvest, treated barley ears were cut, weighed, immediately frozen in liquid nitrogen and stored at −80 °C until further processing.
Fruit were harvested, treated with 100 uL.L-1 ethylene, and assessed as above 1 day, 2 days and 4 days following treatment (Additional file 4).
Lysates from HEK293Ts stably expressing N-terminally tagged 3× FLAG TLR9 and GFP tagged UNC93B1-WT, -H412R, or -Y539A were harvested, treated with EndoH, separated by SDS-PAGE, and visualized by immunoblot with anti-FLAG and anti-GFP antibodies.
The total RNA was extracted by TRIzol after harvesting cells treated with fenofibrate.
Lymph nodes, thymus and spleen were harvested from treated mice, mechanically disrupted, passed through a 40 µm cell strainer (BD Falcon) and rinsed with PBS to remove debris.
Livers and blood were harvested from treated and untreated controls (n = 9 per group) and GSH concentrations, GPx activity, thiobarbituric acid reactive substances (TBARS), 8-isoprostanes and plasma lipid concentrations were measured.
