Technical Reference #1797

Glass Bottom Culture Dishes

Citation in paper containing MatTek reference:
Cells were plated at a concentration of 7.5 105 cells/ml on poly-L-lysine-coated coverslips (MatTek, Ashland, MA).

1797.

Glucocorticoid-mediated Inhibition of Lck Modulates the Pattern of T Cell Receptor-induced Calcium Signals by Down-regulating Inositol 1,4,5-Trishosphate Receptors Michael W. Harr, Yiping Rong, Martin D. Bootman, H. Llewelyn Roderick, Clark W. Distelhorst, Case Western Reserve University, University Hospitals of Cleveland, Babraham Institute, University of Cambridge, Journal of Biological Chemistry, 284(1797), (2009)
Link To Paper

Materials & Methods:
Fura-2AM was purchased from Invitrogen. Dasatinib (BMS-354825, Sprycel) and thapsigargin were purchased from LC laboratories (Woburn, MA). Dexamethasone was purchased from Sigma. Lck and IP3 receptor siRNAs were purchased from Dharmacon (Lafayette, CO). The following antibodies were used in this study: anti-mouse CD3e (145-2C11) and IP3 receptor 3 (BD Biosciences); Lck (3A5) (Santa Cruz Biotechnology, Santa Cruz, CA); phospho-Lck Tyr-394 (Src Tyr-416, 2101) (Cell Signaling Technology, Danvers, MA); -actin (AC-15) (Sigma). Antibody against Type I IP3 receptor was kindly provided by Jan Parys and HumbertDeSmedt (Rbt 03) (KU, Leuven, Belgium) (26). Antibodies for Type I and Type II IP3 receptors were also kindly provided by Richard Wojcikiewicz (CT-1 and CT-2; SUNY, Syracuse, NY) (27, 28).

Microscopic Technique
inverted Zeiss axiovert S100 microscope and 20X Fluor objective

Cell Type(s)
WEHI7.2 cells