RAW264.7 was divided into six treatment groups: control group, LPS (Sigma-Aldrich, St. Louis, MO, USA) group, LPS + Ang-(1–7) (APExBIO, Houston, TX, USA) group, LPS + Ang-(1–7) + A-779 (APExBIO) group, Ang-(1–7) group, and A-779 group. A-779 is the Mas receptor antagonist and was preincubated for 30 minutes before treatment with LPS and Ang-(1–7).
Lipopolysaccharides (lps)
Manufactured by Apexbio
Sourced in United States
The LPS is a high-performance laboratory equipment designed for a wide range of applications. It features precise temperature control, advanced safety mechanisms, and reliable operation. The core function of the LPS is to provide a controlled environment for various experimental and analytical processes.
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Lab products found in correlation
Lipopolysaccharides (lps), by Merck Group (3 mentions)
Ang 1 7, by Apexbio (1 mentions)
A 779, by Apexbio (1 mentions)
Lipopolysaccharides (lps), by Selleck Chemicals (1 mentions)
Bay11 7082, by Apexbio (1 mentions)
Cenicriviroc, by Selleck Chemicals (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Primocin, by InvivoGen (1 mentions)
Kw2449, by MedChemExpress (1 mentions)
Ripa 56, by MedChemExpress (1 mentions)
Fetal bovine plasma, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Gap26, by Apexbio (1 mentions)
5 protocols using lipopolysaccharides (lps)
1
Ang-(1-7) Modulates LPS-Induced Inflammation
2
Acute Inflammation Response in Mice
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The littermate male WT and Lf−/− mice (6–8 weeks) were weighed and injected intraperitoneally with LPS (L9143, Sigma–Aldrich) at a dose of 4 mg/kg to induce acute inflammation response (or with an equal volume of PBS as the control group, each group n = 5). For Cenicriviroc treatment experiment, the mice were intraperitoneally injected with Cenicriviroc (CVC, 10 mg/kg, Selleck) 10 min prior to LPS injection. 4 h later, the mice were weighed again (each group n = 3). Serum was collected and stored at − 80 °C. The mice were injected intraperitoneally with 5 ml of RPMI-1640 after euthanasia, and the peritoneal lavage fluid was collected and centrifuged to obtain the lavage fluid supernatant and peritoneal cells. The WT and Lf−/− MEF cells were stimulated by LPS (1 μg/ml) or PBS for 4 h, and BAY11-7082 (1 μM, APExBIO) is added to inhibit the NF-κB pathway.
3
LPS-Induced Acute Lung Injury Model
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SENP3fl/fl and SENP3 cKO mice were intraperitoneally (i.p.) injected with 30 mg/kg LPS (Sigma‐Aldrich). The blood and plasma were collected at 2, 4 and 8 hours post‐LPS injection. After 8 hours, the left lungs were isolated for histopathologic evaluation, whereas bronchoalveolar lavage fluid (BALF) was collected from the right lungs. JNK inhibitor SP600125 (APEXBIO) was dissolved in dimethyl sulfoxide (DMSO) at 15 mg/mL. Male C57BL/6 mice were i.p. injected with 30mg/kg LPS. After 15 minutes, these mice were i.p. injected with SP600125 (75 mg/kg) or DMSO. Mouse survival from LPS injection following SP600125 treatment or DMSO was monitored every 4 hours for up to 60 hours after LPS injection. The blood and BALF were obtained at 4 hours post‐LPS injection. After 8 hours, the total lungs were isolated for histopathologic evaluation.
4
Murine Macrophage-Like Cell Inflammation and Necroptosis
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The murine macrophage-like cell line RAW 264.7 was obtained from the ATCC. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% (v/v) fetal bovine plasma (Gibco) with 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco) and 100 μg/ml Primocin (In vivoGen) at 37°C in a humid atmosphere with 5% CO2. Then, RAW 264.7 cells were reseeded in 96-well or 6-well culture plates at densities of 1×105 or 1×106 cells/well and used for follow-up experiments. RAW 264.7 cells were stimulated with LPS (1 μg/ml) to establish a cell inflammation model and LPS (1 μg/ml) plus zVAD (20 μM; APExBIO) to establish a necroptosis model, with or without KW2449 (1 μM) or RIPA56 (1 μM; MedChemExpress LLC). The cell supernatant was collected 24 h after drug addition, and the cells were collected at the corresponding time points for detection.
5
Modulating H9c2 Cell Response to LPS
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H9c2 cells were purchased from The Shanghai Institute of Biological Sciences, Chinese Academy of Sciences. This cell line is currently used for screening molecular mechanisms in vitro. H9c2 cells were cultured in 10% FBS (Gibco, Carlsbad, CA, USA) and DMEM (Gibco, Thermo Fisher Scientific, Inc.). The control group did not undergo any processing. APN (PeproTech, Rocky Hill, NJ, United States) was added at three concentrations (0.5, 1 and 2 μg/ml) for 2 h, and 1 μg/ml LPS (Sigma, USA) was then added for 24 h. In addition, the Cx43 inhibitor Gap26 (APExBIO Technology LLC, 0.5 μmol/L, 30 min) was added to the preconditioned cells before LPS treatment. The cells were also pre-treated with LY294002 at a concentration of 10 μM for 1 h and then treated with 2 μg/ml APN for 2 h before treatment with LPS for 24 h.
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