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. 2016 Aug 23;7(39):63466–63487. doi: 10.18632/oncotarget.11525

Figure 5. Integrin β1 is required for motility and FAK is required for activation of PI3K-AKT signaling after MEK suppression.

Figure 5

Figure 5

Figure 5

Figure 5

(A–D) Integrin β1 or β5 were depleted by siRNA transfection in parental, BoM2, BrM2, and LM2 cells prior to incubation with drug. After transfection of siRNA, cells were treated with either DMSO or 25 μM UO126 for additional 48 hr. Control siRNA was used for negative control. Migration was then measured by wound healing assay. (E) Parental, BoM2, BrM2, and LM2 cell lines were transfected for 48 hr with control siRNA, Integrin β1 siRNA, or Integrin β5 siRNA. Cells were then pre-incubated with either DMSO or 25 μM UO126 for additional 48 hr. Cell extracts were immunoblotted to measure the level of phosphorylated AKT. (F) Prior to analysis of p-FAK and p-Pyk2 by western blot, either DMSO or 25 μM UO126 was added and cells were incubated for 48 hr. Cell extracts were collected with 2X lamelli sample buffer and immunoblotted with indicated antibodies (G) Parental and LM2 cells were transfected for 48 hr with control siRNA, FAK siRNA, or Pyk2 siRNA as 50 nM concentration. Cells were then pre-incubated with either DMSO or 25 μM UO126 for additional 48 hr. (H–I) Prior to western blot analysis, Parental, BrM2, BoM2 and LM2 cells were treated in combination with both UO126 (25 μM) and PF573228 (10 μM) for 24 hr.