Fig. 4.

Foxp3 expression was positively correlated with the expression of immunity-associated molecules. a, b A2058 cells were transfected using only Lipofectamin2000 (lipo), control siRNA (pSil-ctrl), or Foxp3 siRNA (pSil-Foxp3). Cells were recovered 48 h before the analysis of Foxp3 expression by RT–PCR (a) and 72 h before Western blotting (b). c–f relative expression of Foxp3, B7-H1, TGF-β1, and TGF-β2 after specific silencing or overexpression of Foxp3. Following transfection with lipo, pSil-ctrl, pSil-Foxp3, pRc-ctrl (control expression vector), or pRc-Foxp3 (Foxp3 expression vector), the mRNA transcription levels of Foxp3 (c), B7-H1, TGF-β1, and TGF-β2 (d) were measured using semiquantitative PCR. Foxp3 and B7-H1 proteins (e) were analyzed using Western blotting, and the TGF-β1 and TGF-β2 concentrations in the culture supernatants (f) were determined by ELISA