Table 1

Expression of HLA-E, -F, and -G Transcripts in Sorted HLA-Ghigh Thymic Epithelial Cells

PrimersLocationPCR fragmentExpression (% of pan-HLA class I)*
bp
HLA-E
 5′-TCCGAGCAAGAATCAAATGATGExon 370041.5 ± 12
 5′-GTGTGAGGAAGGGGGTCATG3′-untranslated
HLA-F
 5′-TATTGGGAGTGGACCACAGGGTACExon 284231.4 ± 10
 5′-GGCACAAGTGCAATTCTGCTAC3′-untranslated
HLA-G
 5′-TTGGGAAGAGGAGACACGGAACACExon 28663.3 ± 2.4
 5′-GGGCTGGTCTCTGCACAAAGAGA3′-untranslated
pan-HLA class I
 5′-TCCCACTCCATGAGGTATTTCExon 2777100
 5′-CACATGGCAGGTGTATCTCTGExon 4
  • Messenger RNA from HLA-Ghigh thymic epithelial cells purified by fluorescence-activated cell sorting, was amplified by PCR using either locus-specific oligonucleotides or pan-HLA primers corresponding to conserved sequences in all class I genes. Each PCR reaction was performed using mRNA from 5 × 103 cells. PCR fragments obtained in the linear range of amplification were separated by agarose gel electrophoresis, stained by ethidium bromide, and analyzed by densitometry. The pixel intensity of each nonclassical HLA transcript is expressed as percentage of that measured for the total HLA transcript, amplified with pan-HLA primers.  

  • *  Data represent mean ± SEM of three independent cell sorting experiments.