ad5null a20 viruses a431 arwyn jones epidermoid cancer n a n a egfr ve cell line  (ATCC)

Journal: Cellular and molecular life sciences : CMLS

Article Title: Tetraspanin-enriched Microdomains Regulate Digitation Junctions

doi: 10.1007/s00018-018-2803-2

Figure Lengend Snippet: A. Role of PI-4 kinase signaling in DJs. PI4P and EWI2/PGRL are colocalized in DJs and memtubs of A431 cells. B. Role of PI-3 kinase signaling in DJs. Du145 cells cultured on glass coverslips were treated with wortmannin (100 nM) in serum-free medium at 37oC for 60 mins before they were fixed, incubated with CD9 mAb, DAPI, and phalloidin, and examined with confocal microscopy. C. Role of MLCK signaling. Du145 cells cultured on glass coverslips were treated with ML-7 (30 μM) in serum-free medium at 37°C for 60 mins before they were fixed, incubated with CD9 or CD44 mAbs, DAPI, and phalloidin, and then examined with confocal microscopy. Quantification of microextrusion density in DJs (mean±S.E, n=3 individual experiments). ***: P<0.001. Scale bars: 10 μm.

Article Snippet: Cells used in this study include human microvascular endothelial cells (HMECs) (CDC, Atlanta, GA), Du145 and LnCap human prostate cancer (ATCC, Manassas, VA), U87 human glioblastoma (ATCC), and A431 human epidermoid cancer cell lines (ATCC), Du145-Mock and -CD82 stable transfectants 18 , 48 , HT1080-Mock and HT1080-CD9 stable transfectants, PC3 human prostate cancer cells (ATCC) in which EWI2/PGRL was transiently silenced with siRNA oligo against the target sequence GUUCUCCUAUGCUGUCUU of EWI2/PGRL or in which control siRNA oligo was transfected 22 , MDCK-GFP:CD82 and -GFP:CD151 stable transfectants expressing the fusion proteins in which eGFP moieties were fused to the N-termini of CD82 and CD151 proteins, respectively, and NIH3T3-EWI2/PGRL transfectant 25 .

Techniques: Cell Culture, Incubation, Confocal Microscopy