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Bacteria were washed after each incubation with 10% porcine serum/PBS and after two final washes, cell pellets were resuspended in 1% paraformaldehyde.
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One hour after incubation, bacteria were washed out and medium was changed with one containing antibiotics (gentamycin 100 µg/mL).
After the bacteria were washed several times with PBS containing 1% mannose, they were incubated together with the villi for one hour at room temperature.
After appropriate dilutions, bacteria were washed twice with phosphate-buffered saline and mice were fed 50 µl of the solution containing 109 CFU with a pipette to the back of the oropharynx after fasting for 16 h.
The morning after that, the bacteria were washed twice with saline and then resuspended in 2 mL of filtered, distilled water.
To begin to assess whether the SapM mutation alters DC-mediated immune responses, BM-DCs were infected with parental M. bovis BCG or with its SapM mutant derivative for 6 h, after which the bacteria were washed away.
After extensive HBSS washing, the extracellular bacteria were washed away.
After 48 hours at 28°C the bacteria were washed three times with saline 0.85%.
After incubation with gentle mixing at 37°C, bacteria were washed extensively with PBS and were added to polylysine-coated plates.
The bacteria were washed and diluted in sterile water.
After incubation, 1 mM phenylmethylsulfonyl fluoride (PMSF; Sigma, St . Louis MO) was added for 10 min, and then bacteria were washed three times in PBS.
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CEO of Professional Science Editing for Scientists @ prosciediting.com