Protocol detail

Immunofluorescence Staining of Cell Markers

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Cells were fixed and stained according to published methods [17 (link)]. Treated cells were fixed and incubated with the following antibodies: anti-E-cadherin (R&D Systems, 1:500), anti-β-catenin (1:500, R&D Systems), anti-Zeb1 (1:200, Abcam), and F-actin (Cell Signaling). Following primary incubation, conjugation with FITC (Sigma Biochemicals), Cy3 (Sigma Biochemicals), or Alexa Fluor 594 (Invitrogen) secondary antibodies was then performed. The cells were imaged with a Zeiss Axiophot microscope. Images were merged and analyzed using NIH Image J software.

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Dosch A.R., Dai X., Gaidarski III A.A., Shi C., Castellanos J.A., VanSaun M.N., Merchant N.B, & Nagathihalli N.S. (2019). Src kinase inhibition restores E-cadherin expression in dasatinib-sensitive pancreatic cancer cells. Oncotarget, 10(10), 1056-1069.

Publication 2019

anti-E-cadherin anti-β-catenin anti-Zeb1 F-actin Alexa Fluor 594 Axiophot microscope

Alexa 594 Antibodies Cadherin Cells F actin Fitc Microscope Stained methods β catenin

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Variable analysis

independent variables

Treatment of cells

dependent variables

Expression/localization of E-cadherin
Expression/localization of β-catenin
Expression/localization of Zeb1
Expression/localization of F-actin

control variables

Cell fixation method
Antibody concentrations
Secondary antibody conjugations
Microscope imaging conditions

controls

No treatment (negative control)

Annotations

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