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Hh-dependent luciferase reporter assay

AC Andrea Calcaterra
VI Valentina Iovine
BB Bruno Botta
DQ Deborah Quaglio
ID Ilaria D’Acquarica
AC Alessia Ciogli
AI Antonia Iazzetti
RA Romina Alfonsi
LS Ludovica Lospinoso Severini
LM Lucia Di Marcotullio
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The luciferase assay was performed in NIH3T3 Shh-Light II cells, stably incorporating a Gli-responsive luciferase reporter and the pRL-TK Renilla (normalisation control). The Shh-Light II cells were cultured in DMEM plus 10% FBS and then treated for 48 h with SAG alone (200 nM, Alexis Biochemicals, Farmingdale, NY) or in combination with GANT61 or GANT61-D.

Luciferase and Renilla activity were assayed with a dual-luciferase assay system according to the manufacturer’s instruction (Biotium Inc., Hayward, CA). Results are expressed as luciferase/Renilla ratios and represent the mean ± SD of three independent experiments, each performed in triplicate.

Copyright and License information:  ©2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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