Technical Reference #1879

1879.

Cell-permeable ceramides act as novel regulators of U937 cell–cell adhesion mediated by CD29; CD98; and CD147 Yong Gyu Lee; Jaehwi Lee; Jae Youl Cho, Kangwon National University, Immunobiology, 215(1879), (2010)

Keywords:
Ceramides; Cell adhesion; CD29 (Beta1-integrins); CD98; CD147

Materials & Methods:
Confocal analysis (Lee et al. 2008) of actin and CD29 co-localization on U937 cells (1 105 cells/ml) or actin cytoskeleton in LPS-treated RAW264.7 cells (1 105 cells/ml) treated with ceramide was performed on glass-bottom culture dishes (MatTek Corp. Ashland MA) using FITC-labeled antibodies to CD29 or CD98 and rhodamine palloidin (actin). Intensity changes in CD29 and CD98 or actin cytoskeleton were imaged by LSCM on a Zeiss LSM 510 META confocal microscope equipped with a Zeiss 37 1C incubation system. Images were analyzed using the Zeiss LSM Image Examiner. HEK293 cells (1 105 cells/ml) treated with C6-ceramide (20 mM) or cytochalasin B (20 mM) for 12 h were photographed to evaluate the reagents’ effect on morphological change.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
RAW264.7 cells