1. a b
10A.zp85WT
10A.zp85S83μ
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 S ζ 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 LY 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.