MARC-145 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, 10566-016, Gibco, NY, USA) supplemented with 10% fetal bovine serum (FBS, 10099141C, Gibco), 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate (B540732, Sangon Biotech, Shanghai, China). PAMs were obtained and cultured as previously described (27 (link)). The recombinant PRRSV-GFP strain (50 (link)) was kindly donated by Professor En-Min Zhou from Northwest A&F University (Yangling, Shaanxi, China). The highly pathogenic PRRSV strain HN07-1 (HP-PRRSV, GenBank accession no. KX766378.1 ) (51 (link)) was kindly donated by Professor Gai-Ping Zhang from Henan Agricultural University (Zhengzhou, China). The less pathogenic PRRSV strain BJ-4 (LP-PRRSV no. AF331831.1 ) was used as described previously (27 (link)). Viruses were propagated and titrated in MARC-145 cells by calculating the median tissue culture infective dose as previously described (27 (link)).
B540732
Manufactured by Sangon
Sourced in China
The B540732 is a laboratory equipment designed for general use in scientific research and analysis. It serves as a tool for performing various experimental and testing procedures. The core function of this product is to provide a reliable and standardized platform for conducting laboratory-based investigations and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lps l2630, by Merck Group (1 mentions)
Recombinant human tgf β1 protein, by R&D Systems (1 mentions)
C11330500bt, by Thermo Fisher Scientific (1 mentions)
Streptomycin sulfate, by Sangon (1 mentions)
Penicillin, by Sangon (1 mentions)
5 protocols using b540732
1
Culturing MARC-145 Cells and Propagating PRRSV Strains
2
Culturing MARC-145 Cells and Propagating PRRSV Strains
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MARC-145 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, 10566-016, Gibco, NY, USA) supplemented with 10% fetal bovine serum (FBS, 10099141C, Gibco), 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate (B540732, Sangon Biotech, Shanghai, China). PAMs were obtained and cultured as previously described (27 (link)). The recombinant PRRSV-GFP strain (50 (link)) was kindly donated by Professor En-Min Zhou from Northwest A&F University (Yangling, Shaanxi, China). The highly pathogenic PRRSV strain HN07-1 (HP-PRRSV, GenBank accession no. KX766378.1 ) (51 (link)) was kindly donated by Professor Gai-Ping Zhang from Henan Agricultural University (Zhengzhou, China). The less pathogenic PRRSV strain BJ-4 (LP-PRRSV no. AF331831.1 ) was used as described previously (27 (link)). Viruses were propagated and titrated in MARC-145 cells by calculating the median tissue culture infective dose as previously described (27 (link)).
3
Modeling Diabetic Cardiomyopathy in H9C2 Cells
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H9C2 rat cardiomyocytes were obtained from the National Infrastructure of Cell Line Resource (1101RAT-PUMc000219). H9C2 were cultured in DMEM medium with 5.5 mmol/L glucose (11885084, Gibco, USA), 10% fetal bovine serum (FBS, 10100147, Gibco, USA) and 100 U/mL penicillin-100 μg/mL streptomycin sulfate (B540732, Sangon, China) at 37° C under 5% CO2. According to the literature (4 (link), 21 (link), 22 (link)), in this research, high glucose was defined as 35 mmol/L was used to intervene H9C2 cardiomyocytes for 24 h to establish a DCM model in vitro study.
H9C2 cells were divided into three groups: (1) CON group: H9C2 cells were cultured with DMEM medium containing 5.5 mmol/L glucose. (2) HG group: H9C2 cells were cultured with DMEM medium containing 35 mmol/L glucose. (3) HG + Cana group: H9C2 cells were cultured with DMEM medium containing 35 mmol/L glucose and a final concentration of 10 μM canagliflozin. After 24 h of treatment, the H9C2 cells in each group were measured for related indexes.
H9C2 cells were divided into three groups: (1) CON group: H9C2 cells were cultured with DMEM medium containing 5.5 mmol/L glucose. (2) HG group: H9C2 cells were cultured with DMEM medium containing 35 mmol/L glucose. (3) HG + Cana group: H9C2 cells were cultured with DMEM medium containing 35 mmol/L glucose and a final concentration of 10 μM canagliflozin. After 24 h of treatment, the H9C2 cells in each group were measured for related indexes.
4
In-vitro cell culture and treatment
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HK-2 cells (the Cell Resource Center, Peking Union Medical College) and HMC cells (FuHeng Biology, Shanghai) were cultured in DMEM/F-12 (C11330500BT, Gibco) and DMEM (01-052-1ACS, BI), respectively, supplemented with 10% FBS (04-001-1ACS, BI) and 1% penicillin/streptomycin (B540732, Sangon Biotech). The primary mouse renal tubular epithelial cells (PRTC cells) were isolated and cultured according to our previous report61 . HK-2 cells were treated with 10 ng/mL recombinant human TGF-β1 protein (R&D system, USA), HMC cells were treated with 15 μg/ml LPS (L2630, Sigma, USA), while PRTC cells were treated with 10 ng/mL recombinant human TGF-β1 protein and 30 mM high glucose (A100188, Sangon Biotech). The concentration of 1,5-AG (BYOC-ALD-070-1g, Omicron Biochrmicals) for 24 h treatments of the HK-2, HMC and PRTC cells was 50 μM. The concentration of SBI-115 (TGR5 antagonist, HY-111534, MedChemExpress, Monmouth Junction, NJ, USA) for 48 h treatment of the PRTC cells was 10 μM.
5
Viral Infection Assays in Cell Lines
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PK-15, Vero, MARC-145, and HeLa cells were grown in monolayers at 37 °C under 5% CO2. All cells were cultured in DMEM (10566–016, GIBCO), supplemented with 10% FBS (10099141C, GIBCO), 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate (B540732, Sangon).
PRV-GFP and PRV-QXX were used as previously described [31 (link),32 (link)]. PRV HN1201 [33 ] was a gift from Professor Ke-Gong Tian (Henan Agricultural University, China). Vesicular stomatitis virus (VSV)-GFP, Newcastle disease virus (NDV)-GFP, influenza virus (H1N1 PR8), and Sendai virus (SeV) [34 (link)] were gifts from Yong-Tao Li (Henan Agricultural University, China). Porcine reproductive and respiratory syndrome virus (PRRSV)-GFP [35 (link)] was a gift from Professor En-Min Zhou (Northwest A&F University, China).
PRV-GFP and PRV-QXX were used as previously described [31 (link),32 (link)]. PRV HN1201 [33 ] was a gift from Professor Ke-Gong Tian (Henan Agricultural University, China). Vesicular stomatitis virus (VSV)-GFP, Newcastle disease virus (NDV)-GFP, influenza virus (H1N1 PR8), and Sendai virus (SeV) [34 (link)] were gifts from Yong-Tao Li (Henan Agricultural University, China). Porcine reproductive and respiratory syndrome virus (PRRSV)-GFP [35 (link)] was a gift from Professor En-Min Zhou (Northwest A&F University, China).
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