Research Article

Effects of PARP-1 Deficiency on Th1 and Th2 Cell Differentiation

Figure 4

PARP-1 deficiency does not affect T-bet mRNA expression. (a) and (b) Naïve CD4 T cells from wild type (WT, filled symbols) and PARP-1KO (KO, unfilled symbols) mice were stimulated with anti-CD3 and anti-CD28 mAbs alone or together with either IL-12 and neutralizing anti-IL-4 mAb (Th1-polarizing conditions) or IL-4 and a neutralizing anti-IFN-γ mAb (Th2-polarizing conditions) or anti-IL4 neutralizing mAb (anti-IL4). T-bet mRNA expression was assessed by reverse transcription and real time PCR after 24 (a) and 72 (b) hours of stimulation. Results, normalized on the housekeeping gene (GAPDH) mRNA level, are referred to the T-bet mRNA level in WT cells stimulated for 24 hrs and expressed as arbitrary units. Values represent means ± S.E. Results were confirmed in two other independent experiments. (c) After 5 days stimulation with anti-CD3 and rCD86 alone (nonpolarizing conditions, upper panels) or together with IL-12 (and anti-IL-4) (Th1-polarizing conditions, lower panels), cells were analyzed by flow cytometry. Representative dot plots are shown. Numbers in dot plots represent means (±S.E.) from three independent experiments. * for KO versus WT cells.