Chemokine mRNA expression in DECs and decidual ST cells. (A) mRNA isolated from in vitro–cultured DECs or ST cells derived from the same donor were analyzed by RNase protection assay using hCK-5 multiprobe set. Data are representative of 3 independent experiments. (B,C) mRNA isolated from 3 different primary cultures of decidual ST cells and 2 different primary cultures of DECs were analyzed by RT-PCR for the expression of CXCL10/IP-10 or CX3CL1/fractalkine. RNA isolated from PMA/ionomycin-activated peripheral blood leukocytes (PBLs) was used as positive control for CXCL10/IP-10 and that from U251 glioblastoma cell line as positive control for CX3CL1/fractalkine (Giovanni Bernardini, Giuseppe Sciumé, A. Soriani, and A. Santoni, unpublished observation, June 2003). MW represents the molecular markers. β-actin and GAPDH are shown as mRNA loading control. These results are representative of 3 independent experiments. (D) mRNA isolated from primary cultures of DECs or ST cells obtained from the same donor was analyzed by real-time quantitative PCR assay for the expression of CXCL10/IP-10, CX3CL1/fractalkine, CCL2/MCP-1, and CXCL12/SDF-1. The relative chemokine amount of each sample was normalized with β-actin and expressed as arbitrary units plus SD. Results obtained from 2 different donors are shown.