Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were performed as described previously (27 (link), 28 (link)).
Briefly, total RNA was isolated from the liver tissues using the Rneasy® Mini Kit (QIAGEN, Hilden, Germany). The RNA was reverse-transcribed using a SuperScript VILO cDNA Synthesis Kit (Thermo Fisher Scientific, CA, USA). qRT-PCR was performed by Taqman master mix (Thermo Fisher Scientific, CA, USA) and a 7900 Sequence Detection System (Applied Biosystems, USA). The specific primer information is listed inTable S1
The antibodies used for the immunoblot are LC3B (2775, Cell Signaling Technology, Beverly, MA, USA) and Rubicon (8465, Cell Signaling Technology, Beverly, MA, USA). For signal normalization, anti-GAPDH antibody was used (MAB374, Millipore, Bedford, MA, USA). Membranes were imaged with the ImageQuant LAS 4000 camera system (GE Healthcare). The band intensity was quantified by Image J software.
Briefly, total RNA was isolated from the liver tissues using the Rneasy® Mini Kit (QIAGEN, Hilden, Germany). The RNA was reverse-transcribed using a SuperScript VILO cDNA Synthesis Kit (Thermo Fisher Scientific, CA, USA). qRT-PCR was performed by Taqman master mix (Thermo Fisher Scientific, CA, USA) and a 7900 Sequence Detection System (Applied Biosystems, USA). The specific primer information is listed in
The antibodies used for the immunoblot are LC3B (2775, Cell Signaling Technology, Beverly, MA, USA) and Rubicon (8465, Cell Signaling Technology, Beverly, MA, USA). For signal normalization, anti-GAPDH antibody was used (MAB374, Millipore, Bedford, MA, USA). Membranes were imaged with the ImageQuant LAS 4000 camera system (GE Healthcare). The band intensity was quantified by Image J software.
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