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. 2003 Jul 7;198(1):153–160. doi: 10.1084/jem.20030633

Figure 2.

Figure 2.

Figure 2.

Figure 2.

Failure of IFN-γ to induce IDO expression and activity in early prediabetic NOD female mice. (A) Functional IDO activity in response to IFN-γ was measured in vitro in terms of the ability to metabolize tryptophan to kynurenine with the use of DCs recovered from NOD female and male mice of two different ages (4 or 8 wk) or control C57BL/6 female mice. Kynurenine levels in supernatants were measured by HPLC, and results are the mean ± SD of triplicate samples in one of three experiments. (B) IDO expression in DCs was assessed by Western blot. DCs were recovered from NOD female or male mice or control DBA/2 females and treated overnight with IFN-γ, and IDO expression was investigated with an IDO-specific antibody. The positive control consisted of IDO-expressing MC24 transfectants and the negative control consisted of mock-transfected MC22 cells. Loading controls (not shown in the figure) consisted of samples reprobed with β-actin-specific antibody. One experiments of three. (C) Stat1 phosphorylation in response to IFN-γ was studied in NOD female and male mice at 4 wk and in age-matched control C57BL/6 female mice. DC lysates were resolved on SDS-PAGE and immunoblotted with anti-phosphoStat1 and then reblotted with anti-Stat1, reblots showing equal loading of Stat1 in each set of lanes. One of three experiments with similar results.