Evaluation of GR protein and phosphorylation state in CEM-C1-15 cells rendered sensitive to Dex by several treatments. (A)CEM-C1-15 cells were pretreated with vehicle control (C), FSK (F), U0126 (U) plus SP600125 (SP), U0126 plus (ip), rapamycin (R), or combinations of these treatments for 5.5 hours before adding Dex (D) for an additional 17 hours. Dex-sensitive CEM-C7-14 cells were treated with vehicle control or Dex for 17 hours, and results are included as a positive control. Equal amounts of protein were added to each lane. The results were evaluated by immunoblot with antibodies against phospho-GR (Ser 211), total GR, and β-actin, n = 2–4 for various combinations of treatments. (B) Densitometric analysis of immunoblot from (A). All bands were normalized to β-actin; bars = arbitrary units of variable/densitometry units of β-actin for each corresponding lane.