Adenoviral and Lentiviral APE1 Expression

The FLAG-tagged coding sequence of APE1 was cloned into pcDNA3.1 mammalian expression plasmid (Invitrogen, Carlsbad, CA USA). The APE1 coding sequence from pcDNA3.1/APE1 plasmid was sub-cloned using Xba I and BamH I restriction sites in the adenoviral shuttle vector pACCMV. The recombinant adenovirus expressing APE1 was generated by co-transfecting HEK-293 cells with the shuttle and backbone adenoviral pJM17 plasmids using a Calcium Phosphate Transfection kit (Applied Biological Materials Inc., Richmond, BC) (34 (link)). Lentivirus particles expressing APE1 shRNA or control shRNA were produced by VectorBuilder Inc. (Santa Clara, CA, USA) and then used to transduce OE33 and CPB cells. Control siRNA (sc-29470) and APE1 siRNA (sc-29470) were obtained from Santa Cruz Biotechnology.

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Bhat A.A., Lu H., Soutto M., Capobianco A., Rai P., Zaika A, & El-Rifai W. (2018). Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1. Oncogene, 37(46), 6011-6024.

Publication 2018

pcDNA3.1 mammalian expression plasmid Calcium Phosphate Transfection kit Control siRNA (sc-29470) APE1 siRNA (sc-29470)

Adenoviral Adenovirus Ape1 Backbone Biological Calcium phosphate Cells Coding sequence Hek 293 cells Lentivirus Mammalian Plasmids Shrna Shuttle vector Sirna Transfection

Corresponding Organization :

Other organizations : University of Miami

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Variable analysis

independent variables

Overexpression of APE1 using a recombinant adenovirus
Knockdown of APE1 using lentivirus particles expressing APE1 shRNA

dependent variables

Not explicitly mentioned

control variables

Use of pcDNA3.1 plasmid as a control for APE1 overexpression
Use of control shRNA lentivirus particles as a control for APE1 knockdown
Use of control siRNA as a control for APE1 siRNA

controls

Positive control: pcDNA3.1/APE1 plasmid for APE1 overexpression
Negative control: Control shRNA lentivirus particles for APE1 knockdown
Negative control: Control siRNA for APE1 siRNA

Annotations

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