Inducible AR-V7 Expression in LNCaP Cells

The cumate-inducible wildtype or mutant AR-V7 expression constructs were individually packaged into lentiviral particles in 293T cells, and the lentivirus particles were used to transduce LNCaP cells as described previously [50 (link)]. At 48 hr after transduction, 1 μg/ml puromycin was added to the medium to start selection, and the selection continued for two weeks to obtain polyclonal colonies of resistant cells. AR-V7 was expressed by exposing the cells to 1x cumate (System Biosciences).

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Roggero C.M., Jin L., Cao S., Sonavane R., Kopplin N.G., Ta H., Ekoue D.N., Witwer M., Ma S., Liu H., Ma T., Gioeli D., Raj G.V, & Dong Y. (2020). A detailed characterization of stepwise activation of the androgen receptor variant 7 in prostate cancer cells. Oncogene, 40(6), 1106-1117.

Publication 2020

cumate

293t cells Lentivirus Puromycin

Corresponding Organization : The University of Texas Southwestern Medical Center

Other organizations : Tulane University, University of Virginia

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Variable analysis

independent variables

Cumate-inducible wildtype or mutant AR-V7 expression constructs

dependent variables

Expression of AR-V7 protein

control variables

Lentiviral transduction of LNCaP cells
Puromycin selection for two weeks to obtain polyclonal colonies of resistant cells

controls

Positive control: Cumate-inducible wildtype or mutant AR-V7 expression constructs to induce AR-V7 expression
Negative control: Not explicitly mentioned

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