Fig 2.

Real-time RT-PCR confirms downregulation of genes involved in the immune system in human NK cells treated with Gal-9. We used PCR to verify the 6 genes shown to be maximally differentially expressed (greater than 7-fold change) in the microarray analysis. (A) We confirmed downregulation of five of these six genes by PCR analysis of NK cells isolated from 4 normal control subjects after 24 h of culture with Gal-9, as performed for the microarray experiment. The five genes confirmed by PCR are involved in the immune response, and all, with the exception of one (CISH), play a stimulatory role. The microarray data are shown in the black bars for comparison to the PCR data shown in the white bars. (B) The time course for each of the six genes is shown. The majority of these genes were significantly downregulated after 2 h in culture with Gal-9. (C) In addition, we tested regulation of genes involved in the cytolytic activity of NK cells (perforin and TRAIL) and found them both to be significantly downregulated after 18 and 24 h of culture with Gal-9. The bars show means ± SEM, and Student's t test was used to test statistical significance.