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Missense mutants were constructed using QuikChange Lightning kit (Stratagene) with HGSNAT-TAP as a matrix (see Table S1 for primers).
Detect the StAR protein using the Chemiluminescence Imaging Western Lightning Kit (PerkinElmer).
Site-directed mutagenesis was performed by using the QuikChange Lightning Kit (Stratagene).
Site-directed mutagenesis was performed using the QuickChange Lightning kit (Stratagene, La Jolla, CA, USA).
Site directed mutagenesis was performed using the QuickChange Lightning kit (Agilent Technologies) on these expression plasmids.
Site directed mutagenesis was performed using the QuickChange Lightning kit (Agilent Technologies, Santa Clara, CA) on the pFL142 plasmid.
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DNA was bisulfite converted using the EZ DNA Methylation-Lightning kit (Zymo Research).
Genomic DNA (500 ng) was bisulfite converted using EZ DNA Methylation-Lightning Kit (Zymo Research).
PCR thermocycling program profile used was as following: 35 cycles of 98º for 10 sec, 64º for 15 sec, and 68º for 10 min. DNA was converted using the EZ DNA Methylation-Lightning kit (Zymo Research, Irvine, CA).
A QuikChange Lightning Mutagenesis kit (Agilent Technologies, CA, USA) with the following primers was used to get the WT and mutant DNA.
Peroxidase-coupled antibodies were detected by enhanced chemiluminescence with the Western Lightning ECL kit (Perkin Elmer).
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