Total RNA was isolated using TRIzolTM (Invitrogen), and cDNA was synthesized using 2 μg of total RNA with SuperScript® III First-Strand Synthesis System, according to the manufacturer’s protocol (Invitrogen). Real-time PCR was performed on LightCycler 480® (Roche), using SYBR Green I Master Mix. The primer set sequences are provided in Supplementary Table S1 .
Total cell lysates were prepared, and western blot analyses were conducted as described previously [32 (link)]. Antibodies are listed inSupplementary Table S2 . Signals were visualized with Western Lighting Plus-ECL Chemiluminescent Reagents (Perkin Elmer, Waltham, MA, USA), and the intensity of the resulting bands was quantified by ImageJ software (NIH, Bethesda, MD, USA).
Total cell lysates were prepared, and western blot analyses were conducted as described previously [32 (link)]. Antibodies are listed in
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