Total RNA was extracted from the cells using TRIzol reagent. One microgram total RNA from each sample was then converted into complementary DNA (cDNA) by reverse transcription [using the PrimeScript™ RT reagent kit (Takara, Dalian, China)]. Quantitative PCR (qPCR) was performed to gauge the mRNA levels using the 7500 Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). The PCR cycling conditions were as follows: 94°C pre-degeneration for 4 min, 94°C degeneration for 30 sec, 60°C annealing for 30 sec, 72°C extension for 2 min, for 35 cycles. The data were analyzed using System SDS software (Applied Biosystems). The expression level of the GAPDH gene was considered as a standard. The relative gene expression levels of interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), fatty acid synthase (Fas), Fas ligand (FasL) and Bcl-2-associated X protein (Bax) were expressed as a percentage of the expression of GAPDH. Respective primers (Table I ) were applied by calculating target gene expression.
System sds software
Manufactured by Thermo Fisher Scientific
Sourced in United States
System SDS software is a laboratory information management system (LIMS) designed to manage safety data sheets (SDS) for chemical compounds and hazardous materials. The software provides a centralized database to store, organize, and retrieve SDS documents, ensuring compliance with regulatory requirements.
Automatically generated - may contain errors
Lab products found in correlation
Abi 7500 real time pcr system, by Thermo Fisher Scientific (2 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Transgene (1 mentions)
Rnaiso plus, by Transgene (1 mentions)
Revertaid kit, by Thermo Fisher Scientific (1 mentions)
Reliaprep rna cell miniprep system, by Promega (1 mentions)
Gotaq qpcr master mix with bryte green, by Promega (1 mentions)
3 protocols using system sds software
1
Quantifying Gene Expression via qPCR
2
Quantitative RT-PCR Analysis of Extracellular Matrix Genes
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Total RNA from HSC-T6 cells was extracted using RNAiso Plus (Transgen Biotech, Beijing, China) following the manufacturer’s protocol, and the purity of the extracted RNA was determined. The primers of COL3A1 [collagen type III alpha 1 chain], COL1A1 (collagen type I alpha 1 chain), α-SMA (α-smooth muscle actin), SDC-4 (syndecan-4), and GAPDH are listed in Table 1 . cDNA was synthesized using a PrimeScript® RT reagent kit according to the manufacturer’s instructions (Transgen Biotech, Beijing, China). For real-time PCR assay, SYBR® Premix Ex TaqTM II (Transgen Biotech, Beijing, China) was used and subjected to quantitative PCR in an ABI 7500 Real Time PCR System, and the data was analyzed using System SDS software (Applied Biosystems, United States).
3
Quantitative Gene Expression Analysis
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Total RNA from cultured cells was extracted using ReliaPrep RNA Cell Miniprep System (Promega, Madison, WI, USA). RNA was reverse-transcribed into complementary DNA using the Revert Aid kit (Thermofisher) and performed at 25 °C for 10 min, 30 min at 50 °C. For quantitative real-time polymerase chain reaction (qPCR), GoTaq qPCR Master Mix with BRYTE green (Promega) was used and the samples were subsequently subjected to qPCR in an ABI 7500 real-time PCR system and analyzed using System SDS software (Applied Biosystems). For analysis according to the comparative Ct (δδCt) method, each Ct value was first normalized to the house-keeping gene Gapdh. Gene-specific primers were produced by Integrated DNA Technologies (Suppl. table 1 ).
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