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10A. z

a. b. 10A. z p85 S83 μ. 10A. z p85 WT. 10A.vec. 10A. z. 10A.vec. 10A z. 10A z .p85 WT. 10A z .p85 S83 μ. E-cadherin. Vimentin. p85. p53. P-Akt. T-Akt. b -actin. 14-3-3 z. c. d. 10A.vec. 10A. z. 10A.vec. 10A. z. -. -. -. -. LY294002. 24hr. 48hr. 24hr. 48hr.

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10A. z

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  1. a b 10A.zp85S83μ 10A.zp85WT 10A.vec 10A.z 10A.vec 10Az 10Az.p85WT 10Az.p85S83μ E-cadherin Vimentin p85 p53 P-Akt T-Akt b-actin 14-3-3z c d 10A.vec 10A.z 10A.vec 10A.z - - - - LY294002 24hr 48hr 24hr 48hr E-cadherin Vimentin p53 P-Akt T-Akt b-actin 14-3-3z Supplementary Figure S7. 14-3-3ζ overexpression induced EMT is independent of PI3K-Akt pathway activation. a. MCF10A.ζ cells were infected with lentiviral pLove constructs expressing untagged p85WT or p85S83μ. Akt activation was assessed by immunoblot for phospho-Akt Ser473 (P-Akt). EMT reversion was assessed by immunoblot for EMT markers E-cadherin (epithelial cell maker) and Vimentin (mesenchymal cell marker). b. Morphology of transfected cells in 2D culture. c. MCF10A.ζ cells were untreated (-) or treated (+) with PI3K inhibitor LY294002 for 24 or 48 hrs. MCF10A.vec cells were used as control. Akt activation was assessed by immunoblot for phospho-Akt Ser473 (P-Akt) to confirm inhibition PI3K. EMT reversion was assessed by immunoblot for EMT markers E-cadherin and Vimentin. d. Morphology of LY29002 treated cells in 2D culture. Scale bars represent 20mm.

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