After 24 hours of infection, supernatants from uninfected, stimulated and infected cells were collected and stored at −80°C until ELISAs were performed.
Cells (1.5 × 105 cells/dish) were starved in 60 mm culture dishes, stimulated for 27 hrs and collected in PBS containing 50 μg/ml propidium iodide, 0.1% (v/v) sodium citrate, 0.1% (v/v) Nonidet P-40.
The neutrophils and supernatants from stimulated neutrophils were collected and used for cell migration and tube formation assays, respectively.
At baseline and at the end of the test period, the plaque index (PI) and gingival index (GI) were determined, and stimulated saliva was collected.
Resting and stimulated saliva were collected to determine flow rate, pH and buffering capacity.
Cells transfected with Rac1 plasmids or stimulated with PDGF were washed and collected in ice-cold PBS following by lysis in IP buffer.
In the morning, saliva was stimulated by chewing Parafilm® and collected into ice-cooled beakers.
Supernatants from synoviocytes stimulated with cytokines were cultured and collected 72 hours after cytokine addition.
We let the subjects chew on a paraffin pellet for 2 min and collected the stimulated saliva.
Both resting and stimulated saliva were collected from the study population between 9 00 to 12 00 noon no earlier than 2 hours after meal.
