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. Author manuscript; available in PMC: 2010 Aug 24.
Published in final edited form as: J Immunol. 2007 Sep 15;179(6):3792–3803. doi: 10.4049/jimmunol.179.6.3792

Figure 7. IFN-α does not promote stable T-bet expression in human CD4+ T cells.

Figure 7

Purified human CD4+/CD45RA+ T cells were activated with plate-bound anti-CD3 + anti-CD28, IL-2, and anti-IL-4, and with either anti-IFNAR2 and anti-IL-12 (“Ctl”), anti-IFNAR2 and IL-12 (“IL-12”), IFN-αA and anti-IL-12 (“IFN-aA”), or with IFN-αA and IL-12 (“IL-12 + IFN-αA”). A, After 72 hours, cells were stained for surface expression of IL-12Rβ2: filled histogram, neutralizing antibodies alone; dashed line, IL-12 + anti-IFNAR2; dotted line, IFN-αA + αIL-12; solid line, IL-12 + IFN-αA. B, Total RNA was isolated from cells harvested 48 hours or 7 days after activation. Analysis of IL-12Rβ2 and T-bet transcript levels was performed by quantitative real-time PCR (qPCR) using the primers listed in Materials and Methods. Transcript levels for each condition were normalized to GAPDH, and the data were further normalized relative to cells activated under neutralizing conditions. C, Whole-cell lysates were prepared from day 7 activated cells and assessed for expression of T-bet protein by Western blotting (upper panel). Blots were stripped and re-probed for Lamin protein expression (lower panel).