Lentiviral Transduction of DTC-1 Cells

The lentiviruses were prepared and used in a similar way as previously described (44 (link)). Briefly, lentiviruses expressing CRE-RLuc or indicated shRNA were prepared from HEK 293T cells by co-transfecting lentiviral expression plasmids along with packaging vectors using the calcium-phosphate method (TakaRa). DTC-1 cells were transduced with lentiviruses expressing CRE-RLuc. After puromycin selection, the cells were used directly for reporter assays.

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Li B.X., David L.L., Davis L.E, & Xiao X. (2022). Protein arginine methyltransferase 5 is essential for oncogene product EWSR1-ATF1-mediated gene transcription in clear cell sarcoma. The Journal of Biological Chemistry, 298(10), 102434.

Publication 2022

calcium-phosphate method

293t cells Assays Calcium phosphate Cells Lentiviruses Plasmids Puromycin Shrna Vectors

Corresponding Organization : Oregon Health & Science University

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Variable analysis

independent variables

Lentiviruses expressing CRE-RLuc or indicated shRNA

dependent variables

Reporter assays

control variables

Lentiviruses were prepared and used in a similar way as previously described (44 (link))
DTC-1 cells were transduced with lentiviruses expressing CRE-RLuc
After puromycin selection, the cells were used directly for reporter assays

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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