Figure 2.

Transcription of c-Myc and Nfkb1 is reduced in Cd28^−/− or Ox40^−/− memory CD8⁺ T cells. Naive Thy1.2 CD8⁺ TCRVβ5⁺ T cells from OT-I, OT-I/Cd28^−/− or OT-I/Ox40^−/− mice were adoptively transferred into Thy1.1 congenic mice infected i.p. with VV-OVA. At day 35, approximately 1 × 10⁶ Thy 1.2⁺ memory T cells from the spleen and LNs were sorted. (a) The scatter plots of PCR array. Total RNA (1 µg) was isolated for the RT² Profiler PCR Array. Pairwise comparison of Wt and Cd28^−/− or Ox40^−/− by scatter plot analysis. Spots associated with individual transcription factor genes were collected and converted into a log₁₀ scale. The central line indicates unchanged gene expression. The dots are allocated to positions that are above or below than the +3 fold or −3 fold line when the differences are greater than threefolds. Data are representative of three independent experiments. (b) Relative expression of c-Myc and Nfkb1 by RT-PCR. Data are represented as the mean ± s.e.m. from three independent experiments (*p < 0.05, **p < 0.01. One-way ANOVA). (c) Protein expression of c-Myc and canonical NF-κB activity detected by immunoblot or p50 ELISA. Similar data were obtained in three experiments (***p < 0.001; two-way ANOVA). In Western blots, β-actin was used as internal control. For Western blots belonging to the same experiment, bands pertaining to different proteins were cropped from the same blot.