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p53 reporter assay

JL Jianshuang Li
DL Di Lu
HD Hong Dou
HL Huadie Liu
KW Kevin Weaver
WW Wenjun Wang
JL Jiada Li
EY Edward T.H. Yeh
BW Bart O. Williams
LZ Ling Zheng
TY Tao Yang
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HEK293 cells (ATCC) were transfected with PG13-Luc (with p53 binding cis-element, 0.5 μg) and Myc-TRIM28, TRIM28K554R, TRIM28K779R, TRIM28K804R, TRIM283KR (0.5 μg) with HA-SUMO3 (0.5 μg) or Flag-SENP6 (1.5 μg) present or absent based on the experimental strategy. Each transfection included Flag-p53 (0.05 μg) and pRL-Renilla (0.05 μg, as control). After incubation for 48 h at 37 °C, the cell lysate was collected for luciferase assay using a Dual-Luciferase kit (Promega). The firefly-luminescence results were normalized to the Renilla luminescence for internal control of transfection efficiency.

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