Repression of the HLA-G gene is reversed by demethylating treatment in several cell lines (A and C). Representative Southern blots of RT-PCR product obtained by amplification with pan-HLA-G G.257F/G.1004R (A) or HLA-G5-specific G.526F/G.i4b (B) primer sets on the JAR, LCL 721.221, Raji, NKL, KG1a, Tera-2, and M8 cell lines, either untreated (U) or treated with increasing concentrations of 5-Aza-dC (1, 10, or 100 μM). GR and GI4F oligonucleotide probes were used to detect all HLA-G transcripts and the HLA-G transcripts that encode soluble HLA-G, respectively. (C) Results of real-time RT-PCR analysis showing relative quantities of HLA-G transcripts in treated (1, 10, or 100 μM 5-Aza-dC) and untreated (U) cell lines, compared with those of JEG-3 (assigned a value of 1).