Confidential Information Regarding Oxymatrine Exposed By Industry Experts
Proteins of interest in liver homogenates were resolved using a 9% or 12% gel and developed using an ECL chemiluminescence system (Amersham, Buckinghamshire, UK). 2.10. RNA isolation and real-time reverse transcription polymerase chain reaction analysis Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA,USA) according to the manufacturer's instructions. To obtain cDNA, total RNA (1?��g) was reverse-transcribed using an oligo(dT)16 primer. The cDNA was amplified using a high capacity cDNA synthesis kit (Bioneer, Daejon, Korea) with Liraglutide clinical trial a thermal cycler (Bio-rad, Hercules, CA, USA). Real-time polymerase chain reaction (PCR) was performed with STEP ONE (Applied Biosystems, Foster City, CA, USA) using a SYBR green premix according to the manufacturer's instructions (Applied Biosystems). Primers were synthesized by Bioneer. The following primer sequences were used: mouse SREBP-1 5��- GAGGCCAAGCTTTGGACCTGG-3�� (sense) and 5��- CCTGCCTTCAGGCTTCTCAGG-3�� (antisense); mouse FAS 5��- ATTGCATCAAGCAAGTGCAG-3�� (sense) and 5��- GAGCCGTCAAACAGGAAGAG-3�� (antisense); mouse ACC 5��- TGAAGGGCTACCTCTAATG-3�� Oxymatrine (sense) and 5��- TCACAACCCAAGAACCAC-3�� (antisense); mouse PPAR�� 5��- CTGCAGAGCAACCATCCAGAT-3�� (sense) and 5��- GCCGAAGGTCCACCATTTT -3�� (antisense); and mouse Sirt1 5��-ATCGGCTACCGAGACAAC-3�� (sense) and 5��- GTCACTAGAGCTGGCGTGT-3�� (antisense). The relative level of PCR products was determined on the basis of the threshold cycle value. Glyceraldehyde-3-phosphate dehydrogenase was used as a reference gene for normalization. Melting curve analysis was done after amplification to verify the accuracy of the amplicon. 2.11. Data analysis One-way analysis of variance was used to assess significant differences among treatment groups. The Newman�CKeuls test was used for comparisons of group means. Statistical analyses were carried out using IBM-SPSS Statistics ver. 21.0 (IBM Corporation, Armonk, NY, USA) for Windows software. Verubecestat in vitro Data represent the mean?��?standard deviation. The criterion for statistical significance was set at p?
