Kinome Profiling of Regulatory T Cells: A Closer Look into a Complex Intracellular Network
Fig 4
Expression levels of marker molecules via qRT-PCR and Western blot.
(A) Treg or Teff were untreated or stimulated with anti-CD3 and anti-CD28 mAb. EGFR, TGFβ-RII, IRF1 and CK2 mRNA expression 0 min (left) and 15 min (right) after stimulation was analyzed and normalized to EF1α and Teff calculated by delta-delta CT Method. Results are representative for eight independent experiments (n = 8, mean ± SEM, *P < .05; **P < .01). (B) Immunoblot analysis of PAK2 in cytoplasmic (left) and nuclear (right) fractions of human Teff and Treg. Cells were unstimulated (0h) or stimulated with anti-CD3 and anti-CD28 mAbs for 24h or 48h. Blotting with actin (left) and laminin (right) antibodies as loading control. One representative experiment out of three is presented. (C) RT-PCR for PAK2 in Teff was performed after stimulation with anti-CD3 and anti-CD28 mAb at 1h and 16h upon activation (n = 3, mean ± SEM, *P < .05).