Figure 7
From: NF-κB/STAT3/PI3K signaling crosstalk in iMycEμ B lymphoma
PI3K inhibition diminishes NF-κB, STAT3 and Myc activity in iMycEμ-1 cells, and reduces their proliferation and survival. (A) Western blot showing the levels of total AKT, PTEN and phosphorylated AKT (S473 and T308) after treatment with LY294002 (LY). α-tubulin was used as a loading control. (B) MTS/PMS assay after treatment with vehicle control, LY, PD98059 (PD), SB203580 (SB), rapamycin (Rap), or AEG 3482 (AEG) at the indicated concentrations. Data were normalized to DMSO-treatment controls, and error bars represent the standard deviation from a representative experiment performed in triplicate. (C) Representative FACS analyses on LY- (open grey histogram) or DMSO- (filled black histogram) treated cells showing an increase in sub-G0/G1 DNA, as assessed by propidium idodide (PI) staining (left panel), and apoptosis as assessed by increases in both Annexin V (middle panel) and activated caspase 3 (right panel) staining. (D) EMSA showing reduced DNA-binding activity of NF-κB, STAT3 and Myc after treatment with LY, but not PD, SB, AEG or Rap. (E) Western blot demonstrating reduced Myc protein levles after inhibition of PI3K; α-tubulin served as a loading control. (F) NF-κB, STAT3 and Myc DNA-binding activity is reduced in a time-dependent manner after PI3K is inhibited with LY. The incubation time with small-molecule inhibitors was 24 hours unless otherwise indicated.