Figure 4.
IL-15 alters HDM2 expression. A, primary NK cells were treated with IL-15 or medium alone for the indicated times. Cell lysates were analyzed by Western blot using a mouse anti-HDM2 antibody. Blots were reprobed with goat anti-actin. B, primary human NK cells were treated with IL-15 and increasing concentrations of nutlin-3 or with lactacystin for 12 h. Cell lysates were analyzed by Western blot using mouse anti-HDM2 and polyclonal anti-Bid antibodies. Blots were reprobed with a goat anti-actin antibody. C, nutlin-3 was added to NK cells at a final concentration of 5 μmol/L and quantitative PCR for Bid expression was done on reverse-transcribed DNA at the indicated times. Fold change in Bid mRNA levels relative to 0 h nutlin-3 addition. Values ± SE were calculated using Relative Quantification Software (Roche Applied Science). Representative of three individual donors.