Effect of TGF-β on IL-12R expression in alloactivated T cells. Six-day (d6) alloactivated T cells were incubated at 1.106/mL in the presence or absence of TGF-β (1 ng/106cells) at 37°C. Total RNA was extracted after 24, 48, and 72 hours of incubation, and transcripts encoding the IL-12Rβ1 and IL-12Rβ2 subunits, L32 and GAPDH, as loading control were quantitated by ribonuclease protection assays as described in Materials and Methods. (A) RPA bands of selected mRNA accumulation from a representative donor. Relative radioactivity values for IL-12Rβ1 and IL-12Rβ2 transcripts (upper row), after normalization against L32 (lower row) values, expressed in arbitrary units for lanes 1 through 7, respectively, were: 37143, 33773, 28275, 30560, 23923, 19729, and 16587. (B) Relative expression of IL-12Rβ1 and IL-12Rβ2 mRNA obtained from four donors. Data are given as mean (± SD) relative radioactivity values (expressed in arbitrary units) for IL-12Rβ1 and IL-12Rβ2 transcripts from four donors after normalization against the corresponding L32 values.
