To establish a model of the progression of liver cirrhosis to hepatocarcinoma, 8-week-old male Axin2Cre;Rosa26EGFP SD rats were injected intraperitoneally with 50 mg/kg diethylinitrosamine (N0258-1G; Sigma-Aldrich, Shanghai, China) twice per week for 8 weeks. The rats were fed a regular diet for an additional 4 weeks. The rats were biopsied to evaluate the presence of liver cirrhosis and autophagy status. Rats with non-autophagic liver cirrhosis were intraperitoneally injected with rapamycin (2 mg/kg, S1039; Selleck, Shanghai, China) twice per week for 4 weeks. Rats with aberrant autophagic liver cirrhosis were then number randomized and injected intravenously with 1 × 109 TU/ml shRNA-vector, shRNA-Atg3, shRNA-Atg7 and shRNA-HGF lentiviruses in transduction enhancement reagent (Envirus; Engreen Biosystem, Beijing, China) twice per week for 4 weeks. In addition, the rats also received chloroquine (50 mg/kg, an inhibitor of the late stage of autophagy; Sigma-Aldrich) weekly, with SP600125 (a JNK inhibitor, 15 mg/kg, S1460; Selleck) or stattic (a STAT3 inhibitor, 2 mg/kg, S7024; Selleck) administered twice per week by intraperitoneal injection for 4 weeks. A group of normal rats or rats with vehicle injection served as the control where appropriate. The rats were killed after an additional 4-week observation after the final treatment. The entire experimental period was 20 weeks.
Envirus
Manufactured by Engreen Biosystem
Sourced in China
The Envirus is a piece of laboratory equipment designed for the detection and analysis of viruses. It utilizes advanced detection technology to identify the presence and characteristics of various viral agents. The core function of the Envirus is to provide accurate and reliable data to support research, diagnostic, and monitoring applications.
Automatically generated - may contain errors
3 protocols using envirus
1
Modeling Liver Cirrhosis to Hepatocarcinoma
2
Knockdown of GPR91 in hPDLCs
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To knock down the expression of GPR91 in hPDLCs, cells were transfected for 24 h with GPR91 shRNA (25 nM; GenePharma, China) or scramble shRNA (25 nM; GenePharma, China) as a negative control with Envirus™ (Engreen Biosystem) according to the manufacturer's protocol. The culture medium was replaced with complete culture medium 24 h later. At least 72 h after transfection, cells were used for subsequent testing. The sequences of the shRNAs used in this study were as follows: control shRNA (shNC), 5′-GGAGTTATGCCAATGGAAACT-3′, and GPR91 gene shRNA, 5′-GGAGTTATGCCAATGGAAACT-3′.
3
Investigating CDK9 Knockdown in THP-1 Cells
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THP-1 cells were cultured in 24-well plates. Lentiviral vectors carrying CDK9 shRNA or not were transfected into cells, with the assistance of Envirus (Engreen Biosystem). The expression of green fluorescence was observed at 96 h post-transfection. RNA and proteins were extracted to examine the expression of CDK9. THP-1 cells were seeded into 96-well plate and 6-well plate for cytotoxicity analysis and real-time PCR analysis, respectively. Cells were divided into four groups. NC group was transfected with non-targeting scramble and stimulated with PBS. NC and P. gingivalis group was transfected with non-targeting scramble and stimulated with P. gingivalis. CDK9 shRNA group was transfected with CDK9 shRNA and stimulated with PBS. CDK9 shRNA and P. gingivalis group was transfected with CDK9 shRNA and stimulated with P. gingivalis. After transfection for 96 hours, cells were stimulated with P. gingivalis and PBS, then 4 hours later, cytotoxicity analysis was detected by LDH assay. RNA was extracted at 2 hours post-stimulation.
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