Luciferase reporter assay

YY Yuhei Yahiro
SM Shingo Maeda
MM Masato Morikawa
DK Daizo Koinuma
GJ Go Jokoji
TI Toshiro Ijuin
SK Setsuro Komiya
RK Ryoichiro Kageyama
KM Kohei Miyazono
NT Noboru Taniguchi
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Luciferase reporter constructs of mouse Atoh8 SBRs, SBR1 (GGCGTC) or SBR2 (GGCGCC), were described previously.32 Mutations in these motifs, GataTC for SBR1 and GaattC for SBR2, were also generated.32 The 6xOSE2 luciferase reporter plasmid was a kind gift from Dr. Toshihisa Komori (Nagasaki University, Japan). The Renilla luciferase reporter vector pGL4.74 (hRluc/TK) (Promega, Madison, WI, USA) was used as an internal control. Cells were seeded in triplicate in 24-well plates and transiently transfected with reporter luciferase plasmids with or without expression vectors. Dual luciferase assays were performed using a GloMax 96 microplate luminometer (Promega).

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