Vol 41 (10) 2011, DOI 10.1002/eji.201141666
TGF-β signaling via Smad4 drives IL-10 production in effector Th1 cells and reduces T-cell trafficking in EAE
David J. Huss, Ryan C. Winger, Gina Mavrikis Cox, Mireia Guerau-de-Arellano, Yuhong Yang, Michael K. Racke, Amy E. Lovett-Racke
In the above article, the method detailing the transfection protocol with siRNA was missing from the Materials and methods. The missing section is provided below with the references following the order of the above article.
Transfection with siRNA in vitro.
All siRNA was purchased from Thermo Fisher Scientific. The IL-10 siRNA sequence was previously verified . Effector Th1 cells were generated from TCR-tg mice and plated in 1.25 ml of EAE media in 6-well plates (7 × 106 cells/well). As previously described , transfection solution was prepared using 200 μl serum-free media, 5 μl Mirus TKO transfection reagent, and 160 μM siRNA, incubated at room temperature for 30 minutes, and added to each well for 18 hours. Cells were then washed with PBS and activated with MBP Ac1–11 (1 μg/ml) alone or in combination with TGF-β (1.0 ng/ml).