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Beyotime protease inhibitor mixture
Protease Inhibitor Mixture, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/protease inhibitor mixture/product/Beyotime
Average 99 stars, based on 18 article reviews
Price from $9.99 to $1999.99
protease inhibitor mixture - by Bioz Stars, 2020-11
99/100 stars

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Centrifugation:

Article Title: Platelet-Derived Exosomal MicroRNA-25-3p Inhibits Coronary Vascular Endothelial Cell Inflammation Through Adam10 via the NF-κB Signaling Pathway in ApoE−/− Mice
Article Snippet: .. After centrifugation at 1,500 × g, the supernatant added with protease and phosphatase inhibitor (P1046, Beyotime Biotechnology Co., Ltd., Shanghai, China) was resuspended with 200 μL PBS and preserved at −80°C for the following experiment ( ). .. Next, a transmission electron microscope (TEM) was used to identify the exosomes.

Radio Immunoprecipitation:

Article Title: Overexpression of NAC1 confers drug resistance via HOXA9 in colorectal carcinoma cells
Article Snippet: .. Western blot analysis Total proteins from tissues and cells were extracted using radioimmunoprecipitation assay buffer containing protease and phosphatase inhibitors (Beyotime Institute of Biotechnology, Haimen, China). .. For western blot analysis, 60 µg of total protein lysates underwent 10% SDS-PAGE and were subsequently transferred onto polyvinylidene difluoridemembranes (Bio-Rad Laboratories, Inc., Hercules, CA, USA) as described previously ( ).

Protease Inhibitor:

Article Title: Chondroprotective Effects of Combination Therapy of Acupotomy and Human Adipose Mesenchymal Stem Cells in Knee Osteoarthritis Rabbits via the GSK3β-Cyclin D1-CDK4/CDK6 Signaling Pathway
Article Snippet: .. Western blot analysisTotal proteins were extracted from the cartilage using ultrasonication in RIPA lysis buffer containing 1% protease inhibitor cocktails (Beyotime Biotechnology, Shanghai, China), and then the protein concentrations were quantified with a bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology, Shanghai, China). .. Equal quantities (50 µg) of proteins were separated by electrophoresis through 12% sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) (PPLYGEN, Beijing, China) gels and then transferred onto 0.45 µm PVDF membranes (Millipore, USA).

Article Title: Suppressing endoplasmic reticulum stress-related autophagy attenuates retinal light injury
Article Snippet: .. Western blot analysisCell, retina and RPE/choroid mixture samples were sonicated in protein lysate buffer (Beyotime, Shanghai, China) containing 1% protease inhibitor cocktails (Beyotime, Shanghai, China). .. A bicinchoninic acid assay was used to measure the protein concentration.

Bicinchoninic Acid Protein Assay:

Article Title: Chondroprotective Effects of Combination Therapy of Acupotomy and Human Adipose Mesenchymal Stem Cells in Knee Osteoarthritis Rabbits via the GSK3β-Cyclin D1-CDK4/CDK6 Signaling Pathway
Article Snippet: .. Western blot analysisTotal proteins were extracted from the cartilage using ultrasonication in RIPA lysis buffer containing 1% protease inhibitor cocktails (Beyotime Biotechnology, Shanghai, China), and then the protein concentrations were quantified with a bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology, Shanghai, China). .. Equal quantities (50 µg) of proteins were separated by electrophoresis through 12% sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) (PPLYGEN, Beijing, China) gels and then transferred onto 0.45 µm PVDF membranes (Millipore, USA).

Western Blot:

Article Title: Overexpression of NAC1 confers drug resistance via HOXA9 in colorectal carcinoma cells
Article Snippet: .. Western blot analysis Total proteins from tissues and cells were extracted using radioimmunoprecipitation assay buffer containing protease and phosphatase inhibitors (Beyotime Institute of Biotechnology, Haimen, China). .. For western blot analysis, 60 µg of total protein lysates underwent 10% SDS-PAGE and were subsequently transferred onto polyvinylidene difluoridemembranes (Bio-Rad Laboratories, Inc., Hercules, CA, USA) as described previously ( ).

Article Title: Chondroprotective Effects of Combination Therapy of Acupotomy and Human Adipose Mesenchymal Stem Cells in Knee Osteoarthritis Rabbits via the GSK3β-Cyclin D1-CDK4/CDK6 Signaling Pathway
Article Snippet: .. Western blot analysisTotal proteins were extracted from the cartilage using ultrasonication in RIPA lysis buffer containing 1% protease inhibitor cocktails (Beyotime Biotechnology, Shanghai, China), and then the protein concentrations were quantified with a bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology, Shanghai, China). .. Equal quantities (50 µg) of proteins were separated by electrophoresis through 12% sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) (PPLYGEN, Beijing, China) gels and then transferred onto 0.45 µm PVDF membranes (Millipore, USA).

Article Title: Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration
Article Snippet: .. Western blotting Proteins were isolated using RIPA lysis buffer with protease inhibitors and phosphatase inhibitors (Beyotime, Nantong, China). .. The proteins were separated by SDS–PAGE gel electrophoresis and transferred to polivinyledene fluoride (PVDF) membranes.

Article Title: Suppressing endoplasmic reticulum stress-related autophagy attenuates retinal light injury
Article Snippet: .. Western blot analysisCell, retina and RPE/choroid mixture samples were sonicated in protein lysate buffer (Beyotime, Shanghai, China) containing 1% protease inhibitor cocktails (Beyotime, Shanghai, China). .. A bicinchoninic acid assay was used to measure the protein concentration.

Real-time Polymerase Chain Reaction:

Article Title: GDNFOS1 knockdown decreases the invasion and viability of glioblastoma cells
Article Snippet: .. CFX96 Touch™ Real-Time PCR Detection system was supplied by (Bio-Rad Laboratories, Inc., and lysis buffer (cat. no. P0013), protease and phosphatase inhibitors (cat. no. P1045), enhanced chemiluminescence solution (cat. no. P0018A), polyvinylidene fluoride (PVDF) membranes (cat. no. FFP24), BCA kit (cat. no. P0012) and film (cat. no. FF057) were all supplied by Beyotime Institute of Biotechnology. .. Primary antibody against GDNF (cat. no. ab176564) and horse-radish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (cat. no. ab205718) were supplied by Abcam.

Incubation:

Article Title: MG132 Attenuates the Replication of Classical Swine Fever Virus in vitro
Article Snippet: .. Moderate amount of cell lysis buffer containing proteasome inhibitors and phosphatase inhibitors (Beyotime, P1045) was added to the cell monolayer and incubated for 15 min on ice. .. Cell lysates were clarified by centrifugation at 12,000 g for 5 min.

Lysis:

Article Title: KLF4 overexpression decreases the viability, invasion and migration of papillary thyroid cancer cells
Article Snippet: .. Tissues were digested and lysed in lysis buffer (cat. no. P0013; Beyotime Institute of Biotechnology) at 4°C with inhibitors of phosphatase and protease (cat. no. P1045; Beyotime Institute of Biotechnology). .. The lysis mixture was centrifuged at 4°C for 10 min at 10,000 × g and the supernatant containing cellular proteins was utilized in the following experiments.

Article Title: Chondroprotective Effects of Combination Therapy of Acupotomy and Human Adipose Mesenchymal Stem Cells in Knee Osteoarthritis Rabbits via the GSK3β-Cyclin D1-CDK4/CDK6 Signaling Pathway
Article Snippet: .. Western blot analysisTotal proteins were extracted from the cartilage using ultrasonication in RIPA lysis buffer containing 1% protease inhibitor cocktails (Beyotime Biotechnology, Shanghai, China), and then the protein concentrations were quantified with a bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology, Shanghai, China). .. Equal quantities (50 µg) of proteins were separated by electrophoresis through 12% sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) (PPLYGEN, Beijing, China) gels and then transferred onto 0.45 µm PVDF membranes (Millipore, USA).

Article Title: Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration
Article Snippet: .. Western blotting Proteins were isolated using RIPA lysis buffer with protease inhibitors and phosphatase inhibitors (Beyotime, Nantong, China). .. The proteins were separated by SDS–PAGE gel electrophoresis and transferred to polivinyledene fluoride (PVDF) membranes.

Article Title: GDNFOS1 knockdown decreases the invasion and viability of glioblastoma cells
Article Snippet: .. CFX96 Touch™ Real-Time PCR Detection system was supplied by (Bio-Rad Laboratories, Inc., and lysis buffer (cat. no. P0013), protease and phosphatase inhibitors (cat. no. P1045), enhanced chemiluminescence solution (cat. no. P0018A), polyvinylidene fluoride (PVDF) membranes (cat. no. FFP24), BCA kit (cat. no. P0012) and film (cat. no. FF057) were all supplied by Beyotime Institute of Biotechnology. .. Primary antibody against GDNF (cat. no. ab176564) and horse-radish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (cat. no. ab205718) were supplied by Abcam.

Article Title: MG132 Attenuates the Replication of Classical Swine Fever Virus in vitro
Article Snippet: .. Moderate amount of cell lysis buffer containing proteasome inhibitors and phosphatase inhibitors (Beyotime, P1045) was added to the cell monolayer and incubated for 15 min on ice. .. Cell lysates were clarified by centrifugation at 12,000 g for 5 min.

Sonication:

Article Title: Suppressing endoplasmic reticulum stress-related autophagy attenuates retinal light injury
Article Snippet: .. Western blot analysisCell, retina and RPE/choroid mixture samples were sonicated in protein lysate buffer (Beyotime, Shanghai, China) containing 1% protease inhibitor cocktails (Beyotime, Shanghai, China). .. A bicinchoninic acid assay was used to measure the protein concentration.

BIA-KA:

Article Title: GDNFOS1 knockdown decreases the invasion and viability of glioblastoma cells
Article Snippet: .. CFX96 Touch™ Real-Time PCR Detection system was supplied by (Bio-Rad Laboratories, Inc., and lysis buffer (cat. no. P0013), protease and phosphatase inhibitors (cat. no. P1045), enhanced chemiluminescence solution (cat. no. P0018A), polyvinylidene fluoride (PVDF) membranes (cat. no. FFP24), BCA kit (cat. no. P0012) and film (cat. no. FF057) were all supplied by Beyotime Institute of Biotechnology. .. Primary antibody against GDNF (cat. no. ab176564) and horse-radish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (cat. no. ab205718) were supplied by Abcam.

Isolation:

Article Title: Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration
Article Snippet: .. Western blotting Proteins were isolated using RIPA lysis buffer with protease inhibitors and phosphatase inhibitors (Beyotime, Nantong, China). .. The proteins were separated by SDS–PAGE gel electrophoresis and transferred to polivinyledene fluoride (PVDF) membranes.

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    Beyotime a 172 glioma cell lines
    MYBL2 is required for T-96-induced cell growth and proliferation inhibition. a MYBL2 expression was analysed via Western blot assays after cells were treated with T-96. Cells were treated with the indicated concentration of T-96 for the indicated times. In the concentration gradient group, cells were treated with different concentrations of T-96 (2, 5 and 10 μM) for two days, and DMSO was used as the control; in the time gradient group, cells were treated with 10 μM T-96 for different times (1, 2 and 3 days), and DMSO was used as the control. b , c The effect of DMSO or T-96 on the viability of MYBL2/EGFP-overexpressing LN-229 and <t>A-172</t> cells. d Representative image of BrdU staining of MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days; scale bar = 100 μm. e Quantification of BrdU-positive staining rates in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days. f Cell cycle distribution was analysed in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or T-96 for 2 days. g Western blot assays were performed to evaluate proteins related to the cell cycle and DNA replication in MYBL2-overexpressing or EGFP-overexpressing LN-229 or A-172 cells after treatment with DMSO or T-96 for 2 days. Tubulin was used as the control. h Densitometry of Western blot in the panel g . All data were analysed using unpaired Student’s t -tests and are shown as the means ± SD. * p
    A 172 Glioma Cell Lines, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a 172 glioma cell lines/product/Beyotime
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    a 172 glioma cell lines - by Bioz Stars, 2020-11
    92/100 stars
      Buy from Supplier

    88
    Beyotime l6 drgs
    Effects of 2-Hz EA on p-PKC ε expressions in L4–L6 DRGs of PDN rats after EA treatment for 7 days. a Representative bright-field micrographs showing p-PKC ε -immunoreactive neurons in L4, L5, and <t>L6</t> DRGs of rats. Scale bar = 100 μm. b Statistical analysis of L4, L5, and L6 DRG p-PKC ε -immunoreactive neurons. Scale bar = 100 μm. Data were presented as mean ± SD, n = 3 per group. ∗∗ P
    L6 Drgs, supplied by Beyotime, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/l6 drgs/product/Beyotime
    Average 88 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    l6 drgs - by Bioz Stars, 2020-11
    88/100 stars
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    MYBL2 is required for T-96-induced cell growth and proliferation inhibition. a MYBL2 expression was analysed via Western blot assays after cells were treated with T-96. Cells were treated with the indicated concentration of T-96 for the indicated times. In the concentration gradient group, cells were treated with different concentrations of T-96 (2, 5 and 10 μM) for two days, and DMSO was used as the control; in the time gradient group, cells were treated with 10 μM T-96 for different times (1, 2 and 3 days), and DMSO was used as the control. b , c The effect of DMSO or T-96 on the viability of MYBL2/EGFP-overexpressing LN-229 and A-172 cells. d Representative image of BrdU staining of MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days; scale bar = 100 μm. e Quantification of BrdU-positive staining rates in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days. f Cell cycle distribution was analysed in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or T-96 for 2 days. g Western blot assays were performed to evaluate proteins related to the cell cycle and DNA replication in MYBL2-overexpressing or EGFP-overexpressing LN-229 or A-172 cells after treatment with DMSO or T-96 for 2 days. Tubulin was used as the control. h Densitometry of Western blot in the panel g . All data were analysed using unpaired Student’s t -tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: MYBL2 is required for T-96-induced cell growth and proliferation inhibition. a MYBL2 expression was analysed via Western blot assays after cells were treated with T-96. Cells were treated with the indicated concentration of T-96 for the indicated times. In the concentration gradient group, cells were treated with different concentrations of T-96 (2, 5 and 10 μM) for two days, and DMSO was used as the control; in the time gradient group, cells were treated with 10 μM T-96 for different times (1, 2 and 3 days), and DMSO was used as the control. b , c The effect of DMSO or T-96 on the viability of MYBL2/EGFP-overexpressing LN-229 and A-172 cells. d Representative image of BrdU staining of MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days; scale bar = 100 μm. e Quantification of BrdU-positive staining rates in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days. f Cell cycle distribution was analysed in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or T-96 for 2 days. g Western blot assays were performed to evaluate proteins related to the cell cycle and DNA replication in MYBL2-overexpressing or EGFP-overexpressing LN-229 or A-172 cells after treatment with DMSO or T-96 for 2 days. Tubulin was used as the control. h Densitometry of Western blot in the panel g . All data were analysed using unpaired Student’s t -tests and are shown as the means ± SD. * p

    Article Snippet: LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Inhibition, Expressing, Western Blot, Concentration Assay, BrdU Staining, Staining

    The miR-30e-5p antagomir (Antago) blocked the effects induced by T-96 in glioma cells. a Quantity real-time PCR (qRT-PCR) assays were performed to evaluate the expression of miR-30e-5p after treatment of LN-229 and A-172 cells with DMSO or 10 μM T-96 for the indicated time. b The expression of miR-30e-5p after T-96 treatment or T-96 and the miR-30e-5p antagomir treatment for 2 days. DMSO was used as the control. c LN-229 and A-172 cells were treated with DMSO, 10 μM T-96, the miR-30e-5p antagomir, or T-96 and the miR-30e-5p antagomir for 2 days, and the cell viability was evaluated with MTT assays. d LN-229 and A-172 cells were treated with DMSO, the miR-30e-5p antagomir, 10 μM T-96 or T-96 and the miR-30e-5p antagomir for 2 days, and cell cycle was analysed via flow cytometry. e Western blot assays were used to detect the expression of MYBL2, CDK4, CDK6 and cyclin D1 after treatment with DMSO, the miR-30e-5p antagomir, 10 μM T-96, or T-96 and the miR-30e-5p antagomir for 2 days. f Densitometry of Western blot in the panel e . All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: The miR-30e-5p antagomir (Antago) blocked the effects induced by T-96 in glioma cells. a Quantity real-time PCR (qRT-PCR) assays were performed to evaluate the expression of miR-30e-5p after treatment of LN-229 and A-172 cells with DMSO or 10 μM T-96 for the indicated time. b The expression of miR-30e-5p after T-96 treatment or T-96 and the miR-30e-5p antagomir treatment for 2 days. DMSO was used as the control. c LN-229 and A-172 cells were treated with DMSO, 10 μM T-96, the miR-30e-5p antagomir, or T-96 and the miR-30e-5p antagomir for 2 days, and the cell viability was evaluated with MTT assays. d LN-229 and A-172 cells were treated with DMSO, the miR-30e-5p antagomir, 10 μM T-96 or T-96 and the miR-30e-5p antagomir for 2 days, and cell cycle was analysed via flow cytometry. e Western blot assays were used to detect the expression of MYBL2, CDK4, CDK6 and cyclin D1 after treatment with DMSO, the miR-30e-5p antagomir, 10 μM T-96, or T-96 and the miR-30e-5p antagomir for 2 days. f Densitometry of Western blot in the panel e . All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Article Snippet: LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, MTT Assay, Flow Cytometry, Cytometry, Western Blot

    T-96 inhibited cell growth and proliferation in human glioma cells. a Concentration- and time-dependent effects of T-96 on glioma cells, including LN-229, U-87, A-172, U-251 and U-118 cells. Left, glioma cells were treated with different concentrations of T-96 for 2 days; right, cells were treated with 10 μM T-96 for different times. Survival was evaluated using MTT assays, and the data are presented as the means ± SD, n = 3). b Images of LN-229 and U-87 cells positive for BrdU staining after treatment with DMSO or the indicated concentration of T-96. Scale bar = 100 μm. The histogram demonstrates the results of the quantification of the number of BrdU-positive cells in LN-229 and U-87 cells. c Soft agar assays were used to investigate the colony formation abilities of LN-229 and U-87 cells after treatment with DMSO or the indicated concentrations of T-96. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: T-96 inhibited cell growth and proliferation in human glioma cells. a Concentration- and time-dependent effects of T-96 on glioma cells, including LN-229, U-87, A-172, U-251 and U-118 cells. Left, glioma cells were treated with different concentrations of T-96 for 2 days; right, cells were treated with 10 μM T-96 for different times. Survival was evaluated using MTT assays, and the data are presented as the means ± SD, n = 3). b Images of LN-229 and U-87 cells positive for BrdU staining after treatment with DMSO or the indicated concentration of T-96. Scale bar = 100 μm. The histogram demonstrates the results of the quantification of the number of BrdU-positive cells in LN-229 and U-87 cells. c Soft agar assays were used to investigate the colony formation abilities of LN-229 and U-87 cells after treatment with DMSO or the indicated concentrations of T-96. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Article Snippet: LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Concentration Assay, MTT Assay, BrdU Staining

    T-96 inhibited cell migration and invasion in human glioma cells. a Transwell migration assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. b The statistical analysis is presented in histograms, and the migration rates were normalised by the proliferation rate. c Transwell invasion assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. d The statistical analysis is presented in histograms, and the invasion rates were normalised by the proliferation rate. e The expression of E-cadherin and vimentin were analysed by Western blot. Tubulin was used as the control. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. ** p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: T-96 inhibited cell migration and invasion in human glioma cells. a Transwell migration assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. b The statistical analysis is presented in histograms, and the migration rates were normalised by the proliferation rate. c Transwell invasion assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. d The statistical analysis is presented in histograms, and the invasion rates were normalised by the proliferation rate. e The expression of E-cadherin and vimentin were analysed by Western blot. Tubulin was used as the control. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. ** p

    Article Snippet: LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Migration, Expressing, Western Blot

    MYBL2 is required for T-96-induced cell growth and proliferation inhibition. a MYBL2 expression was analysed via Western blot assays after cells were treated with T-96. Cells were treated with the indicated concentration of T-96 for the indicated times. In the concentration gradient group, cells were treated with different concentrations of T-96 (2, 5 and 10 μM) for two days, and DMSO was used as the control; in the time gradient group, cells were treated with 10 μM T-96 for different times (1, 2 and 3 days), and DMSO was used as the control. b , c The effect of DMSO or T-96 on the viability of MYBL2/EGFP-overexpressing LN-229 and A-172 cells. d Representative image of BrdU staining of MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days; scale bar = 100 μm. e Quantification of BrdU-positive staining rates in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days. f Cell cycle distribution was analysed in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or T-96 for 2 days. g Western blot assays were performed to evaluate proteins related to the cell cycle and DNA replication in MYBL2-overexpressing or EGFP-overexpressing LN-229 or A-172 cells after treatment with DMSO or T-96 for 2 days. Tubulin was used as the control. h Densitometry of Western blot in the panel g . All data were analysed using unpaired Student’s t -tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: MYBL2 is required for T-96-induced cell growth and proliferation inhibition. a MYBL2 expression was analysed via Western blot assays after cells were treated with T-96. Cells were treated with the indicated concentration of T-96 for the indicated times. In the concentration gradient group, cells were treated with different concentrations of T-96 (2, 5 and 10 μM) for two days, and DMSO was used as the control; in the time gradient group, cells were treated with 10 μM T-96 for different times (1, 2 and 3 days), and DMSO was used as the control. b , c The effect of DMSO or T-96 on the viability of MYBL2/EGFP-overexpressing LN-229 and A-172 cells. d Representative image of BrdU staining of MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days; scale bar = 100 μm. e Quantification of BrdU-positive staining rates in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or 10 μM T-96 for 2 days. f Cell cycle distribution was analysed in MYBL2-overexpressing or EGFP-overexpressing LN-229 and A-172 cells after treatment with DMSO or T-96 for 2 days. g Western blot assays were performed to evaluate proteins related to the cell cycle and DNA replication in MYBL2-overexpressing or EGFP-overexpressing LN-229 or A-172 cells after treatment with DMSO or T-96 for 2 days. Tubulin was used as the control. h Densitometry of Western blot in the panel g . All data were analysed using unpaired Student’s t -tests and are shown as the means ± SD. * p

    Article Snippet: Western blot analysis LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Inhibition, Expressing, Western Blot, Concentration Assay, BrdU Staining, Staining

    The miR-30e-5p antagomir (Antago) blocked the effects induced by T-96 in glioma cells. a Quantity real-time PCR (qRT-PCR) assays were performed to evaluate the expression of miR-30e-5p after treatment of LN-229 and A-172 cells with DMSO or 10 μM T-96 for the indicated time. b The expression of miR-30e-5p after T-96 treatment or T-96 and the miR-30e-5p antagomir treatment for 2 days. DMSO was used as the control. c LN-229 and A-172 cells were treated with DMSO, 10 μM T-96, the miR-30e-5p antagomir, or T-96 and the miR-30e-5p antagomir for 2 days, and the cell viability was evaluated with MTT assays. d LN-229 and A-172 cells were treated with DMSO, the miR-30e-5p antagomir, 10 μM T-96 or T-96 and the miR-30e-5p antagomir for 2 days, and cell cycle was analysed via flow cytometry. e Western blot assays were used to detect the expression of MYBL2, CDK4, CDK6 and cyclin D1 after treatment with DMSO, the miR-30e-5p antagomir, 10 μM T-96, or T-96 and the miR-30e-5p antagomir for 2 days. f Densitometry of Western blot in the panel e . All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: The miR-30e-5p antagomir (Antago) blocked the effects induced by T-96 in glioma cells. a Quantity real-time PCR (qRT-PCR) assays were performed to evaluate the expression of miR-30e-5p after treatment of LN-229 and A-172 cells with DMSO or 10 μM T-96 for the indicated time. b The expression of miR-30e-5p after T-96 treatment or T-96 and the miR-30e-5p antagomir treatment for 2 days. DMSO was used as the control. c LN-229 and A-172 cells were treated with DMSO, 10 μM T-96, the miR-30e-5p antagomir, or T-96 and the miR-30e-5p antagomir for 2 days, and the cell viability was evaluated with MTT assays. d LN-229 and A-172 cells were treated with DMSO, the miR-30e-5p antagomir, 10 μM T-96 or T-96 and the miR-30e-5p antagomir for 2 days, and cell cycle was analysed via flow cytometry. e Western blot assays were used to detect the expression of MYBL2, CDK4, CDK6 and cyclin D1 after treatment with DMSO, the miR-30e-5p antagomir, 10 μM T-96, or T-96 and the miR-30e-5p antagomir for 2 days. f Densitometry of Western blot in the panel e . All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Article Snippet: Western blot analysis LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, MTT Assay, Flow Cytometry, Cytometry, Western Blot

    T-96 inhibited cell growth and proliferation in human glioma cells. a Concentration- and time-dependent effects of T-96 on glioma cells, including LN-229, U-87, A-172, U-251 and U-118 cells. Left, glioma cells were treated with different concentrations of T-96 for 2 days; right, cells were treated with 10 μM T-96 for different times. Survival was evaluated using MTT assays, and the data are presented as the means ± SD, n = 3). b Images of LN-229 and U-87 cells positive for BrdU staining after treatment with DMSO or the indicated concentration of T-96. Scale bar = 100 μm. The histogram demonstrates the results of the quantification of the number of BrdU-positive cells in LN-229 and U-87 cells. c Soft agar assays were used to investigate the colony formation abilities of LN-229 and U-87 cells after treatment with DMSO or the indicated concentrations of T-96. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: T-96 inhibited cell growth and proliferation in human glioma cells. a Concentration- and time-dependent effects of T-96 on glioma cells, including LN-229, U-87, A-172, U-251 and U-118 cells. Left, glioma cells were treated with different concentrations of T-96 for 2 days; right, cells were treated with 10 μM T-96 for different times. Survival was evaluated using MTT assays, and the data are presented as the means ± SD, n = 3). b Images of LN-229 and U-87 cells positive for BrdU staining after treatment with DMSO or the indicated concentration of T-96. Scale bar = 100 μm. The histogram demonstrates the results of the quantification of the number of BrdU-positive cells in LN-229 and U-87 cells. c Soft agar assays were used to investigate the colony formation abilities of LN-229 and U-87 cells after treatment with DMSO or the indicated concentrations of T-96. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. * p

    Article Snippet: Western blot analysis LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Concentration Assay, MTT Assay, BrdU Staining

    T-96 inhibited cell migration and invasion in human glioma cells. a Transwell migration assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. b The statistical analysis is presented in histograms, and the migration rates were normalised by the proliferation rate. c Transwell invasion assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. d The statistical analysis is presented in histograms, and the invasion rates were normalised by the proliferation rate. e The expression of E-cadherin and vimentin were analysed by Western blot. Tubulin was used as the control. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. ** p

    Journal: Cell Death & Disease

    Article Title: Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis

    doi: 10.1038/s41419-018-1086-8

    Figure Lengend Snippet: T-96 inhibited cell migration and invasion in human glioma cells. a Transwell migration assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. b The statistical analysis is presented in histograms, and the migration rates were normalised by the proliferation rate. c Transwell invasion assays of U-87 and A-172 cells treated with DMSO or 10 μM T-96 for 1 day. d The statistical analysis is presented in histograms, and the invasion rates were normalised by the proliferation rate. e The expression of E-cadherin and vimentin were analysed by Western blot. Tubulin was used as the control. All data were analysed using unpaired Student’s t-tests and are shown as the means ± SD. ** p

    Article Snippet: Western blot analysis LN-229, U-87 and A-172 glioma cell lines were harvested, washed with ice-cold PBS buffer and then suspended in RIPA lysis buffer (Beyotime, China) with phosphatase inhibitors (Sigma Aldrich, St. Louis, MO, USA) and complete protease inhibitor cocktail (Roche).

    Techniques: Migration, Expressing, Western Blot

    Effects of 2-Hz EA on p-PKC ε expressions in L4–L6 DRGs of PDN rats after EA treatment for 7 days. a Representative bright-field micrographs showing p-PKC ε -immunoreactive neurons in L4, L5, and L6 DRGs of rats. Scale bar = 100 μm. b Statistical analysis of L4, L5, and L6 DRG p-PKC ε -immunoreactive neurons. Scale bar = 100 μm. Data were presented as mean ± SD, n = 3 per group. ∗∗ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of 2-Hz EA on p-PKC ε expressions in L4–L6 DRGs of PDN rats after EA treatment for 7 days. a Representative bright-field micrographs showing p-PKC ε -immunoreactive neurons in L4, L5, and L6 DRGs of rats. Scale bar = 100 μm. b Statistical analysis of L4, L5, and L6 DRG p-PKC ε -immunoreactive neurons. Scale bar = 100 μm. Data were presented as mean ± SD, n = 3 per group. ∗∗ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques:

    Effects of intraperitoneal injection of PMA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats subject to 2-Hz EA after EA treatment for 7 days. PMA was intraperitoneally injected (0.2 ng/μl, 100 μl) into the ventral surface of hind paw 10 min prior to EA treatment. Rats in the PDN + EA + PBS group received the same dose of PBS buffer as a control. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the PDN + EA + PBS group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. △ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of intraperitoneal injection of PMA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats subject to 2-Hz EA after EA treatment for 7 days. PMA was intraperitoneally injected (0.2 ng/μl, 100 μl) into the ventral surface of hind paw 10 min prior to EA treatment. Rats in the PDN + EA + PBS group received the same dose of PBS buffer as a control. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the PDN + EA + PBS group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. △ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques: Injection, Western Blot

    Effects of 2-Hz EA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats after EA treatment for 7 days. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the control group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. ∗∗ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of 2-Hz EA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats after EA treatment for 7 days. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the control group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. ∗∗ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques: Western Blot

    Effects of 2-Hz EA on p-PKC ε expressions in L4–L6 DRGs of PDN rats after EA treatment for 7 days. a Representative bright-field micrographs showing p-PKC ε -immunoreactive neurons in L4, L5, and L6 DRGs of rats. Scale bar = 100 μm. b Statistical analysis of L4, L5, and L6 DRG p-PKC ε -immunoreactive neurons. Scale bar = 100 μm. Data were presented as mean ± SD, n = 3 per group. ∗∗ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of 2-Hz EA on p-PKC ε expressions in L4–L6 DRGs of PDN rats after EA treatment for 7 days. a Representative bright-field micrographs showing p-PKC ε -immunoreactive neurons in L4, L5, and L6 DRGs of rats. Scale bar = 100 μm. b Statistical analysis of L4, L5, and L6 DRG p-PKC ε -immunoreactive neurons. Scale bar = 100 μm. Data were presented as mean ± SD, n = 3 per group. ∗∗ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques:

    Effects of intraperitoneal injection of PMA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats subject to 2-Hz EA after EA treatment for 7 days. PMA was intraperitoneally injected (0.2 ng/μl, 100 μl) into the ventral surface of hind paw 10 min prior to EA treatment. Rats in the PDN + EA + PBS group received the same dose of PBS buffer as a control. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the PDN + EA + PBS group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. △ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of intraperitoneal injection of PMA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats subject to 2-Hz EA after EA treatment for 7 days. PMA was intraperitoneally injected (0.2 ng/μl, 100 μl) into the ventral surface of hind paw 10 min prior to EA treatment. Rats in the PDN + EA + PBS group received the same dose of PBS buffer as a control. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the PDN + EA + PBS group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. △ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques: Injection, Western Blot

    Effects of 2-Hz EA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats after EA treatment for 7 days. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the control group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. ∗∗ P

    Journal: Purinergic Signalling

    Article Title: Suppressing PKC-dependent membrane P2X3 receptor upregulation in dorsal root ganglia mediated electroacupuncture analgesia in rat painful diabetic neuropathy

    doi: 10.1007/s11302-018-9617-4

    Figure Lengend Snippet: Effects of 2-Hz EA on total and plasma membrane protein levels of P2X3 receptor in L4, L5, and L6 DRGs of PDN rats after EA treatment for 7 days. a , b Representative Western blot protein and c relative amounts of total and plasma membrane P2X3 receptor in L4 DRGs. d , e Representative Western blot protein and f relative amounts of total and plasma membrane P2X3 receptor in L5 DRGs. g , h Representative Western blot protein and i relative amounts of total and plasma membrane P2X3 receptor in L6 DRGs. Results were expressed as relative fold changes as compared to the control group after normalization to GAPDH. Data were presented as mean ± SD of three independent experiments, n = 5 per group. ∗∗ P

    Article Snippet: Total protein was collected as follows: bilateral L4, L5, and L6 DRGs were sonicated on ice in RIPA Lysis Buffer (Beyotime, China) with an addition of protease inhibitor cocktail (Sangon Biotech, China), centrifuged at 10,000× rpm for 10 min at 4 °C, and then the supernatants were collected.

    Techniques: Western Blot