Pcdna3.1 hmgb1

Manufactured by GenePharma

Sourced in China

PcDNA3.1-HMGB1 is a plasmid vector designed for the expression of the High Mobility Group Box 1 (HMGB1) protein in mammalian cells. It contains the necessary genetic elements for efficient expression and selection of the HMGB1 gene.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Jsh 23, by Abcam (1 mentions) Beas 2b, by National Collection of Authenticated Cell Cultures (1 mentions) Atg5 sirna, by GenePharma (1 mentions) Mtor sirna, by GenePharma (1 mentions) Pbabe puro mek dd, by Addgene (1 mentions) Lv2 pglv u6 puro, by GenePharma (1 mentions) Empty pcdna3.1 plasmid, by GenePharma (1 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions) Hmgb1 sirna, by GenePharma (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Control sirna, by Sangon (1 mentions) Fluoview 2000, by Olympus (1 mentions)

5 protocols using pcdna3.1 hmgb1

NSCLC Cell Line Modulation by miR-449a and HMGB1

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Three human NSCLC cell lines (A549, H1299, and H460) and a normal human lung epithelial cell line (BEAS-2B) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, P.R. China). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Los Angeles, CA, USA) containing 10% fetal bovine serum (FBS; HyClone) at 37°C with 5% CO₂.
The miR-449a mimic, pcDNA3.1-HMGB1, and their corresponding negative control (miR-NC, blank vector) were purchased from GenePharma (Shanghai, P.R. China). The miRNA or HMGB1 transfection was performed using Lipofectamine™ 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.
Additionally, cells without transfection were treated with a NF-κB inhibitor JSH-23 (Abcam, Cambridge, MA, USA) at a concentration of 40 μM.

Wu D., Liu J., Chen J., He H., Ma H, & Lv X. (2019). miR-449a Suppresses Tumor Growth, Migration, and Invasion in Non-Small Cell Lung Cancer by Targeting a HMGB1-Mediated NF-κB Signaling Pathway. Oncology Research, 27(2), 227-235.

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Autophagy Regulation via HMGB1

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The GFP-tagged LC3 cDNA expression construct was a gift from Dr. Noboru Mizushima (Tokyo Medical and Dental University, Tokyo, Japan). P-Babe-Puro-MEK-DD, which constitutively expresses activated MEK1-DD (S218D/S222D), was purchased from Addgene Inc. (Cambridge, MA). Transfections with pcDNA3.1-HMGB1, HMGB1 shRNA, Atg5 siRNA, mTOR siRNA, PI3K-III siRNA and MEK siRNA (all obtained from GenePharma, Shanghai, China) were performed using Lipofectamine 2000 (Invitrogen, USA), according to the manufacturer’s protocol.

Pan B., Chen D., Huang J., Wang R., Feng B., Song H, & Chen L. (2014). HMGB1-mediated autophagy promotes docetaxel resistance in human lung adenocarcinoma. Molecular Cancer, 13, 165.

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Investigating miR-505 and HMGB1 in Renal Cancer

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Selected human renal cancer cells Caki‐1 and 769P were transfected with constructed lentiviral vectors (Lv2‐pGLV‐u6‐puro; GenePharma, Shanghai, China). The empty lentiviral vector was taken as a control. Based on the protocols, cells were infected with lentiviruses and screened with puromycin. Caki‐1 were transfected with miR‐505 mimics while 769P were dealt with miR‐505 inhibitor, NC and anti‐NC were used as negative control.
In addition, selected cell lines were transfected with HMGB1 small interfering RNA (siRNA; GenePharma), negative control siRNA (GenePharma), pcDNA3.1‐HMGB1 (GenePharma), empty pcDNA3.1 plasmid (GenePharma). For plasmid and siRNA transfections, cells were transfected with Lipofectamine 3000 (Thermo Fisher Scientific, Shanghai, China) based on the protocols.

Zhong B., Qin Z., Zhou H., Yang F., Wei K., Jiang X, & Jia R. (2019). microRNA‐505 negatively regulates HMGB1 to suppress cell proliferation in renal cell carcinoma. Journal of Cellular Physiology, 234(9), 15025-15034.

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Etanercept modulates psoriatic keratinocyte proliferation

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HaCaT cells were starved in a serum‐free medium for 12 h before treatment and transfection. Next, HaCaT cells were exposed to different doses of etanercept (0, 5, 10, 20, 40, and 80 μg/ml; Shanghai Sunshine Guojian Pharmaceutical Co., Ltd., China) for 24 h at 37˚C in 5% CO₂. To further examine the effects of etanercept in HaCaT cells, we constructed hyperproliferative psoriatic keratinocytes using LPS.³² HaCaT cells were divided into Control (without treatment), LPS (HaCaT cells at a density of 1 × 10⁵ cells/ml were treated with 10 μg/ml LPS for 24 h), LPS+5 μg/ml Etanercept (HaCaT cells at a density of 1 × 10⁵ cells/ml were treated with 5 μg/ml etanercept for 6 h before LPS treatment), LPS + 40 μg/ml Etanercept (HaCaT cells at a density of 1 × 10⁵ cells/ml were treated with 40 μg/ml etanercept for 6 h before LPS treatment), LPS+Etanercept+si‐HMGB1 (HaCaT cells at a density of 1 × 10⁵ cells/ml were transfected with siRNA‐HMGB1 and treated with 40 μg/ml etanercept for 6 h before LPS treatment), and LPS+Etanercept+HMGB1 (HaCaT cells at a density of 1 × 10⁵ cells/ml were transfected with pcDNA3.1‐HMGB1 and treated with 40 μg/ml etanercept for 6 h before LPS treatment). The plasmid pcDNA3.1‐HMGB1 and siRNA‐HMGB1 were purchased from GenePharma (Shanghai, China). Lipofectamine™ 3000 reagent (Invitrogen, Carlsbad, CA, USA) was used for transfection.

Li S., Li G., Li X., Wu F, & Li L. (2023). Etanercept ameliorates psoriasis progression through regulating high mobility group box 1 pathway. Skin Research and Technology, 29(4), e13329.

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HMGB1 Knockdown Regulates Autophagy in Cells

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Cells were transfected with HMGB siRNA (Sense: GGAAGUUUCUACUGUAUAGTT; Antisense: CUAUACAGUAGAAACUUCCTT)or control siRNA (Sense: UUCUCCGAACGUGUCACGUTT; Antisense: ACGUGACACGUUCGGAGAATT) (Sangon Biotech, Shanghai, China) using lipofectamine2000 (Invitrogen, USA) and Opti-MEM (Invitrogen, USA) according to the manufacturer's protocol, then transfected with adenoviral vectors carrying GFP-LC3for 24 h, treated with GEM for indicated time, fixed in 4% formaldehyde for 10 min, wash with PBS, stained with DAPI for 10 min. The samples were examined under a confocal laser scanning microscope (Olympus Fluoview 2000). The GFP-LC3 puncta in each cell were quantified by Image J software. Transfections with pcDNA3.1 control or pcDNA3.1 HMGB1 (GenePharma, shanghai, China) were performed as mentioned above.

Yin H., Yang X., Gu W., Liu Y., Li X., Huang X., Zhu X., Tao Y., Gou X, & He W. (2017). HMGB1-mediated autophagy attenuates gemcitabine-induced apoptosis in bladder cancer cells involving JNK and ERK activation. Oncotarget, 8(42), 71642-71656.

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