For transfection, cells were seeded at 1.5×105/well in a 6-well plate one day before transfection, and transfected with annealed siRNA using Lipofectamine 2000 (Invitrogen) in a procedure recommended by the manufacturer.
A procedure recommended by OpGen was followed.
CORT levels were measured with a commercially available ELISA kit (Cayman Chemical, MI) using the procedure recommended by the manufacturer.
To determine if any samples harbored the HPV virus, DNA was isolated from the saliva sample using a minimum of 3.5 x 10 cells using the GenomicPrep DNA isolation kit (Amersham Biosciences: Buckinghamshire, United Kingdom), using the procedure recommended by the manufacturer as previously described [ 12, 42, 78, 79].
To determine if any samples harbored the HPV virus, DNA was isolated from the saliva sample using a minimum of 3.5 × 105 cells and the GenomicPrep DNA isolation kit (Amersham Biosciences: Buckinghamshire, UK), using the procedure recommended by the manufacturer, as previously described [ 26, 27, 32].
Approximately 2×106 cells were transfected (Effectene, Qiagen) with either myc- or HA-tagged FraWT, FraΔP1, FraΔP2, and FraΔP3 using the procedure recommended by the manufacturer.
Fatty acid methyl esters (FAME) were identified using the procedure recommended by Microbial Identification System (MIDI, Sherlock Microbial Identification System Version 4.0, MIS Operating Manual, March 2001, Newark, DE).
using the procedure recommended by the manufacturer.
The date of death was determined using the procedure recommended by the GPRD.
The Greek version of the CIS-R was translated and back-translated using the procedure recommended by the World Health Organization http://www.who.int/substance_abuse/research_tools/translation/en/index.html.html
