BCPAP DasRes Cell Genetic Manipulation

BCPAP DasRes cells were transduced with pBabe-hygro or pBabe-c-Src-Dasatinib-Resistant-T3381 retrovirus (Addgene plasmid 26980) and selected with hygromycin (9 (link)). shRNAs targeting p130Cas (Sigma mission TRCN0000115984, TRCN0000115985) or a scrambled control (Sigma mission pLKO.1-puro SHC016) were transduced and selected with puromycin. BCPAP DasRes cells were transfected a gene pool of 5 different siRNAs targeting c-Jun (siJUN) or nontargeting (NT) siRNA (5 nM each) using a final concentration of 0.5% Dharmafect I transfection reagent, according to manufacturerâ€™s protocols Dharmacon (Broomfield, CO).

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Kessler B.E., Mishall K.M., Kellett M.D., Clark E.G., Pugazhenthi U., Pozdeyev N., Kim J., Tan A.C, & Schweppe R.E. (2018). Resistance to Src inhibition alters the BRAF-mutant tumor secretome to promote an invasive phenotype and therapeutic escape through a FAK>p130Cas>c-Jun signaling axis. Oncogene, 38(14), 2565-2579.

Publication 2018

pBabe-hygro pBabe-c-Src-Dasatinib-Resistant-T3381 retrovirus TRCN0000115984 pLKO.1-puro SHC016 Dharmafect I transfection reagent

A gene C jun Cells Dasatinib Hygromycin P130cas Plasmid Puromycin Retrovirus Shrnas Sirna Transfection

Corresponding Organization :

Other organizations : University of Colorado Denver, University of Colorado Cancer Center

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Variable analysis

independent variables

Transduction with pBabe-hygro or pBabe-c-Src-Dasatinib-Resistant-T3381 retrovirus
Transduction with shRNAs targeting p130Cas or a scrambled control
Transfection with a gene pool of 5 different siRNAs targeting c-Jun (siJUN) or nontargeting (NT) siRNA

dependent variables

Not explicitly mentioned

control variables

Selection with hygromycin
Selection with puromycin
Transfection with 0.5% Dharmafect I transfection reagent

controls

Positive control: pBabe-hygro retrovirus transduction
Negative control: Scrambled shRNA control

Annotations

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