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The suggested protocol for treating cells with poly(dA:dT) can be found below:
Poly(dA:dT) Treatment Protocol (5 μg/mL final concentration)
Reagents:
- Poly(deoxyadenylic-thymidylic) acid sodium salt (CST product: Poly(dA:dT) Sodium Salt #47945)
- FuGENE 6 Transfection Reagent from FuGENE
Procedure:
24 hours prior to transfection:- Plate cells in two 15 cm plates with 21 mL media with serum (no antibiotic)
On transfection day:
- Prepare two falcon tubes per plate to be transfected:
- For mock: One tube labeled “FuGENE” and one tube labeled “Mock”
- For poly(dA:dT): One tube labeled “FuGENE” and one tube labeled “Poly(dA:dT)”
- Add 4.5 mL of 1X serum-free medium to ea…
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ZEB2 mRNA is alternatively spliced and there are post-translational modifications (PTMs), such as ubiquitination and sumoylation, that may be the cause of the larger, 210kDa band. The potential modification sites on the ZEB2 protein can be found at https://www.phosphosite.org/proteinAction.action?id=15497&showAllSites=true. Please also see the UniProt page for human ZEB2 for more sequence and protein information (UniProt: O60315, https://www.uniprot.org/uniprot/O60315).