Figure 3

HLA-G expression in PG NSCs. (A) HLA-G-specific RT-PCR analysis of human K562 CML cells, PBMCs, fetal brain (19 wks of gestation), N and PG hESCs and NSCs. Samples were normalized to the housekeeper gene RPL30. Relative gene expression levels were determined using the 2^−ΔΔCt method; HLA-G gene expression in human JEG-3 cells was set to 1. **p < 0.001; n = 3. (B) Western blot analysis to determine HLA-G protein levels in PG and N NSCs (antibody: MEM-G/1). GAPDH was used as loading control (left panel). Densitometric analysis of three independent Western blots was performed using ImageJ. HLA-G protein levels were normalized to GAPDH. *p < 0.05 (right panel). (C) Flow cytometry of untreated (upper panel) or 24-h EDTA-treated (1 mmol/L) (lower panel) NSC cultures. Cells were stained with the HLA-G-specific antibody MEM-G/9. Histograms show specific fluorescence signal for HLA-G (black lines) compared with isotype control (filled gray). Specific fluorescence indexes (SFI: specific geometric median/geometric median of unspecific control) are indicated in the upper right.