Realstar fast sybr qpcr mix kit

To harvest hippocampus tissues, the rats were placed in clean, transparent, and sealed containers connected to a CO₂ Tanker, until they could not move. Then, the rats were extracted and sacrificed immediately by cervical disassociation, one by one. Hippocampus tissues were lysed using the Trizol reagent for the extraction of total RNAs, followed by transforming isolated RNAs into cDNAs with a commercial kit (MedChemExpress, Monmouth Junction, NJ, USA). After conducting the PCR reaction using the 2×RealStar Fast SYBR qPCR Mix kit (GenStar, China), the 2^−ΔΔCt method was utilized for the determination of gene expressions. The primer sequences are listed in Table 1.

Total cellular RNA was extracted with TRIzol reagent (Invitrogen)and cDNA synthesis was performed with HiScript^® II Q RT SuperMix (Vazyme). The relative mRNA levels were quantified via real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) using the 2 × RealStar Fast SYBR qPCR Mix kit (GenStar) on the 7,500 real-time PCR system (Applied Biosystems). β-Actin was used as the reference gene. Primer sequences used in this study were listed in Table 1.

All reagents are commercially sourced. Sources for basic reagents are provided in the supplemental information. Solarbio Technology Co., Ltd. provided the DEAE-Sepharose Fast Flow (Beijing, China). Sephacryl S200HR was purchased from Sigma-Aldrich Company (Darmstadt, Germany). CCK-8 Cell Counting Kit was purchased from Vazyme Biotech Co., Ltd. (Nanjing, China). GenStar Biosolutions Co., Ltd. (Beijing, China) provided 2× Real Star Fast SYBR qPCR Mix kit. Applied Biological Materials Inc (British Columbia, VIC, Canada) provided All-In-One 5× RT MasterMix. Cell Signaling Technology (Boston, MA, USA) provided the rabbit mAbs for β-actin, phospho-NF-κB p65, HO-1, NQO1, and anti-rabbit IgG HRP-linked antibody. PRRS virus nucleocapsid protein mAb was purchased from GeneTex (SAN Antonio, TX, USA). Millipore (Waltham, MA, USA) provided the chemiluminescent HRT substrate. Both the DAPI staining solution and FITC-labeled goat anti-rabbit IgG (H+L) were bought from Beyotime Biotechnology Co., Ltd. in Shanghai, China.