CRTh2 promoter activity
CRTh2 pro-Luc, a luciferase reporter construct containing an 1836
base pair fragment of the CRTH2 promoter, was transiently
co-transfected along with a minimal constitutively active thymidine
kinase renilla control plasmid (pRL-TK) into CCRF-CEM cells as described
[35]. Cells were treated with DEX in
the presence or absence of PPT (22 hours), pelleted, resuspended in
lysis buffer (250 µl), subjected to freeze/thaw and 10 µL of sample
analyzed using Dual-Luciferase Reporter Assay. Protein concentrations
were measured and results expressed as relative luciferase units (RLU),
normalized for transfection efficiency and protein concentration.