Lymph node stromal cells: Control siRNA treated vs. Eif4g3 siRNA treated

Gene expression in the top, light and heavy polysome fractions of Eif4g3 siRNA treated lymph node stromal cells (LNSCs) compared to control-sIRNA treated samples This study was performed to examine whether the expression of certain genes in LNSCs is regulated by the translation factor, eukaryotic translation initiation factor 4 gamma 3 (Eif4g3). LNSCs were grown in 10 cm plates, allowed to reach ≥ 80% confluency and then transfected with 400 pmol of control or Eif4g3 siRNA using Lipofectamine 2000. 7 plates were transfected with control or Eif4g3 siRNA. Cells were pooled, lysed and fractionated on a 10% to 60% continuous sucrose gradient. Fractions were collected and pooled into the top, light and heavy polysome fractions. RNA was extracted from these samples and used for microarrays on the Agilent Whole Mouse Genome Microarray Kit, 4×44K 2-color arrays.