PcDNA-XIST (OV-XIST group), sh-XIST (KD-XIST group), miR-124–3p mimic, miR-124–3p inhibitor, sh-IRF1 and the corresponding negative control were synthesized by Genepharma (Shanghai, China). The above-mentioned plasmids were separately transfected into BV2 cells through Lipofectamine 3000 (Invitrogen, CA, USA). After 48 h, real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) or Western blot assay was utilized to assess the efficiency of transfection.
Sh xist
Manufactured by GenePharma
Sourced in China
Sh-XIST is a laboratory equipment product designed for scientific research. It functions as a tool for the investigation of gene expression and regulation. The product specifications and core capabilities are provided without interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Sh nc, by GenePharma (2 mentions)
Pcdna xist, by GenePharma (1 mentions)
Sh irf1, by GenePharma (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Mir 124 3p mimic, by GenePharma (1 mentions)
Mir 124 3p inhibitor, by GenePharma (1 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000, by Merck Group (1 mentions)
Si malat1, by GenePharma (1 mentions)
Si nc, by GenePharma (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
5 protocols using sh xist
1
Transfection of BV2 Cells with XIST, miR-124-3p, and IRF1 Modulators
2
Plasmid-Mediated IL-6 Expression and RNA Interference
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Plasmid DNA-encoding IL-6 was constructed by inserting the cDNA clone of IL-6 into the pcDNA3.1 vector (Life Technologies, CA, USA). The short hairpin RNA against XIST (sh-XIST) and its negative control (shNC) were purchased from GenePharma (Shanghai, China). And miR-let-7c mimics/inhibitor and mimics/inhibitor NC were obtained from Sigma-Aldrich (CA, USA). For in vitro transfection, cells were transfected using Lipofectamine™ 3000 (Sigma-Aldrich, MO, USA) according to the manufacturer's instructions.
3
MALAT1, MDM4, and XIST Modulation in NSCLC
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Small interfering RNA‐(siRNA) targeting MALAT1 (si‐MALAT1), siRNA‐targeting MDM4 (si‐MDM4) and corresponding negative control (si‐NC), lentiviral particles harboring shRNA targeting XIST (sh‐XIST), and nontarget oligonucleotide sequence (sh‐NC) were structured by Genepharma. The MALAT1 and MDM4 overexpression vectors (MALAT1 and MDM4) and negative control pGL3 empty vector (pcDNA), as well as miR‐185‐5p mimics (miR‐185‐5p) and mimics negative control (miR‐NC) were obtained from Biomics Biotechnologies. All above plasmids or oligonucleotides were transduced into NSCLC cells using Lipofectamine 3000 reagent (Invitrogen) according to the user guide.
4
Spinal Cord Injury Modeling in Mice
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Mice were anesthetized and exposed the T9-11 spinous process and lamina with the T10 lamina as the center. The T10 spinous process and lamina were removed. An impactor with a diameter of 1.5 mm and a weight of 3 g was used to hit the exposed spinal cord, resulting in contusion of moderate severity. Successful modeling was marked by contusion at the site of injury, convulsions in both lower extremities, and spasmodic tail motion. In the sham group, only T10 laminectomy was performed without impingement. One week before SCI modeling, lentivirus-encapsulated pcDNA-XIST (OV-XIST group) and sh-XIST (KD-XIST group) (synthesized by Shanghai Genepharma, China) were injected into the spinal cord of the mice, respectively.
5
Sepsis Induction in Rat Model
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Male Sprague-Dawley rats were acquired from Shanghai SLAC laboratory Animal Co., Ltd.
The polymicrobial sepsis was induced with cecal ligation and puncture (CLP) in rats. Briefly, rats were anesthetized with a gas mixture containing 3% isoflurane and 40% oxygen. A midline surgical incision was performed. The cecum was exposed and ligated by a 3-0 silk suture. The cecum was punctured twice with an 18-gauge needle. Then, the abdomen was closed in layers with 5-0 sutures. For the sham-operated animals, the cecum was mobilized in the absence of CLP. Lentiviral particles were administered through tail veins 1 week before CLP according to a previous study [16] . The rats were randomly assigned to four experimental groups: Sham group (n = 20), CLP group (n = 20), CLP + sh-NC group (n = 20), and CLP + sh-XIST group (n = 20). The rats in CLP + sh-NC and CLP + sh-XIST groups were administered with lentiviral particles containing sh-NC or sh-XIST (1 × 10 9 TU/mL, GenePharma Co., Ltd) through tail veins in a volume of 150 μL. All animal experiments were conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals and approved by the Ethics Committee of Henan Provincial People's Hospital.
The polymicrobial sepsis was induced with cecal ligation and puncture (CLP) in rats. Briefly, rats were anesthetized with a gas mixture containing 3% isoflurane and 40% oxygen. A midline surgical incision was performed. The cecum was exposed and ligated by a 3-0 silk suture. The cecum was punctured twice with an 18-gauge needle. Then, the abdomen was closed in layers with 5-0 sutures. For the sham-operated animals, the cecum was mobilized in the absence of CLP. Lentiviral particles were administered through tail veins 1 week before CLP according to a previous study [16] . The rats were randomly assigned to four experimental groups: Sham group (n = 20), CLP group (n = 20), CLP + sh-NC group (n = 20), and CLP + sh-XIST group (n = 20). The rats in CLP + sh-NC and CLP + sh-XIST groups were administered with lentiviral particles containing sh-NC or sh-XIST (1 × 10 9 TU/mL, GenePharma Co., Ltd) through tail veins in a volume of 150 μL. All animal experiments were conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals and approved by the Ethics Committee of Henan Provincial People's Hospital.
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