HEK293 cells were seeded in clear-bottomed white 96-well plates and transfected in triplicate. For the SV40 assay, cells were transfected with 12 ng of pGL3-promoter firefly luciferase reporter construct (Promega), 5 ng of pRL-TK Renilla luciferase normalization control (Promega), and 16 ng of YFP-FOXP2 (wild-type or K674R or R553H mutant) or YFP control construct. For the SRPX2 assay, cells were transfected with 4.3 ng of SRPX2 luciferase reporter construct, 5 ng of pGL4.74 Renilla luciferase normalization control (Promega), and 45 ng of YFP-FOXP2 (wild-type or K674R or R553H mutant) or YFP control construct. After 48 h, luciferase activity was measured in a TECAN F200PRO microplate reader using the Dual-Luciferase Reporter Assay system (Promega).
Estruch S.B., Graham S.A., Deriziotis P, & Fisher S.E. (2016). The language-related transcription factor FOXP2 is post-translationally modified with small ubiquitin-like modifiers. Scientific Reports, 6, 20911.
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