Non-polarized and polarized T84 cells were pretreated with or without the ezrin inhibitor NSC668394 (20 μM) for 1 h and incubated with GC with or without the inhibitor from the top chamber for 6 h, and then lysed by RIPA buffer [0.1% Triton x100, 0.5% deoxycholate, 0.1% SDS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 2 mM EDTA, 1 mM Na3VO4, 50 mM NaF, 10 mM Na2PO4, and proteinase inhibitor cocktail (Sigma-Aldrich, St. Louis, MO)]. Lysates were resolved using SDS-PAGE gels (BioRad) and analyzed by Western blot. Blots were stained for ezrin (Cell Signaling Technology) and phosphorylated ezrin at T567 (Abcam), stripped and reprobed with anti-β-tubulin antibody (Sigma). Blots were imaged using a Fujifilm LAS-3000 (Fujifilm Medical Systems). Each data point represents individual transwells. Two transwells from two to three independent experiments were quantified.
Phosphorylated ezrin at t567
Manufactured by Abcam
Phosphorylated ezrin at T567 is a protein that functions as a crosslinker between the cell membrane and the actin cytoskeleton. It plays a role in the regulation of cell shape, motility, and signaling.
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Lab products found in correlation
2 protocols using phosphorylated ezrin at t567
1
Ezrin Inhibition in T84 Cells
2
Ezrin Inhibition and GC Exposure
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Non-polarized and polarized T84 cells were pretreated with or without the ezrin inhibitor NSC668394 (20 μM) for 1 h and incubated with GC with or without the inhibitor from the top chamber for 6 h, and then lysed by RIPA buffer [0.1% Triton x100, 0.5% deoxycholate, 0.1% SDS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 2 mM EDTA, 1 mM Na3VO4, 50 mM NaF, 10 mM Na2PO4, and proteinase inhibitor cocktail (Sigma-Aldrich, St. Louis, MO)].
Lysates were resolved using SDS-PAGE gels (BioRad) and analyzed by Western blot. Blots were stained for ezrin (Cell Signaling Technology) and phosphorylated ezrin at T567 (Abcam), stripped and reprobed with anti-β-tubulin antibody (Sigma). Blots were imaged using a Fujifilm LAS-3000 (Fujifilm Medical Systems). Each data point represents individual transwells. Two transwells from two to three independent experiments were quantified.
Lysates were resolved using SDS-PAGE gels (BioRad) and analyzed by Western blot. Blots were stained for ezrin (Cell Signaling Technology) and phosphorylated ezrin at T567 (Abcam), stripped and reprobed with anti-β-tubulin antibody (Sigma). Blots were imaged using a Fujifilm LAS-3000 (Fujifilm Medical Systems). Each data point represents individual transwells. Two transwells from two to three independent experiments were quantified.
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