Technical Reference #1901

1901.

Identification of a Novel Human MD-2 Splice Variant That Negatively Regulates Lipopolysaccharide-Induced TLR4 Signaling Pearl Gray; Kathrin S. Michelsen; Cherilyn M. Sirois; Emily Lowe; Kenichi Shimada; Timothy R. Crother; Shuang Chen; Costantinos Brikos; Yonca Bulut; Eicke Latz; David Underhill; and Moshe Arditi, Cedars-Sinai Medical Center, Journal of Immunology, 184(1901), (2010)
Link To Paper

Materials & Methods:
Confocal imaging TLR4-mCitrine cells were cultured on glass-bottom confocal dishes (MatTek Ashland MA) coated with collagen. After 24 h of culture cells were transfected with up to 300 ng plasmid encoding myc-tagged MD-2 or MD-2s using GeneJuice lipofection reagent (Novagen) and cultured for ∼24 h. Cells were stained with anti-myc Alexa-647 Ab (AbD Serotec) in culture medium at 4°C for 20 min. Cells were gently washed three times with PBS and the culture medium was replaced. Cells were subsequently imaged at room temperature using an SP2 AOBS confocal laser scanning microscope (Leica Microsystems Deerfield IL) running LCS software (Leica Microsystems).

Microscopic Technique
Confocal Microscopy

Cell Type(s)
TLR4-mCitrine cells