lentiviral packaging Search Results


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  • 99
    Thermo Fisher virapower lentivirus packaging mix
    Virapower Lentivirus Packaging Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore mission lentivaral packaging mix
    Chronic GCN5L1 KD attenuated mitochondrial mass and protein levels via autophagic degradation. (A) Representative immunoblot of mitochondrial protein acetylation and the accumulation of autophagy mediators in isolated mitochondria following chronic <t>lentiviral</t>
    Mission Lentivaral Packaging Mix, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mission lentivaral packaging mix/product/Millipore
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    91
    Biosettia lentiviral packaging
    Chronic GCN5L1 KD attenuated mitochondrial mass and protein levels via autophagic degradation. (A) Representative immunoblot of mitochondrial protein acetylation and the accumulation of autophagy mediators in isolated mitochondria following chronic <t>lentiviral</t>
    Lentiviral Packaging, supplied by Biosettia, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lentiviral packaging/product/Biosettia
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    90
    TaKaRa lentiviral packaging
    SynNotch CD19-inducible ROR1 CAR-T cells are unable to rescue toxicity to ROR1 + normal tissues in the presence of circulating and bone marrow-resident CD19 + ROR1 + Raji tumors. ( A) Map of <t>lentiviral</t> constructs encoding CD19-Gal4VP64 synNotch Receptor, UAS inducible R11 ROR1 CAR, and constitutive R11 ROR1 CAR. (B) Raji tumor bioluminescence over time in NSG mice irradiated 250 R and treated with the indicated primary human T cells. n=6–7 per group. Two-way ANOVA with Tukey post-test (UT vs. ROR1 CAR-T: Day 13, 20, p
    Lentiviral Packaging, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lentiviral packaging/product/TaKaRa
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    88
    Thermo Fisher lentivirus packaging mix
    Intein splicing kinetics. ( A ) Autoradiogram after SDS/PAGE and anti-HA immunoprecipitation of lysates from GFP-RecA-HY <t>lentivirus-infected</t> E36 cells pretreated with epoxomicin (5 μM, 2 h) subjected to [ 35 S]-Met/Cys pulse label followed by a “cold”
    Lentivirus Packaging Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 474 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Shanghai Generay Biotech lentiviral packaging
    Intein splicing kinetics. ( A ) Autoradiogram after SDS/PAGE and anti-HA immunoprecipitation of lysates from GFP-RecA-HY <t>lentivirus-infected</t> E36 cells pretreated with epoxomicin (5 μM, 2 h) subjected to [ 35 S]-Met/Cys pulse label followed by a “cold”
    Lentiviral Packaging, supplied by Shanghai Generay Biotech, used in various techniques. Bioz Stars score: 94/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cyagen Biosciences lentiviruses packaging
    Intein splicing kinetics. ( A ) Autoradiogram after SDS/PAGE and anti-HA immunoprecipitation of lysates from GFP-RecA-HY <t>lentivirus-infected</t> E36 cells pretreated with epoxomicin (5 μM, 2 h) subjected to [ 35 S]-Met/Cys pulse label followed by a “cold”
    Lentiviruses Packaging, supplied by Cyagen Biosciences, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore shrna containing lentiviral particles
    Upregulation of core necroptosis components in the spinal cord of symptomatic Tg SOD1 G93A mice. Lumbar spinal cords, from 12- and 15-week-old Tg SOD1 G93A , WT SOD1, and NTg mice, were isolated and processed for mRNA and protein (RIPA or urea extraction) expression of RIPK1, RIPK3, <t>MLKL,</t> and p-MLKL. A , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA from 12-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk3 in Tg SOD1 G93A compared to Tg SOD1 WT ( p = 0.0021) and NTg ( p = 0.0076) at 12 weeks. B , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA in spinal cords of 15-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk1 ( p = 0.0009, vs Tg SOD1 WT ; p = 0.0020, vs NTg) and Ripk3 ( p = 0.0115; vs Tg SOD1 WT , p = 0.0067; vs NTg) but not for Mlkl in Tg SOD1 G93A compared to Tg SOD1 WT and NTg mice at 15 weeks. C , Western blotting (RIPA) for RIPK1 in spinal cord of NTg, Tg SOD1 WT , and Tg SOD1 G93A 15-week-old mice. β-ACTIN, GAPDH: loading control. Specificity of the RIPK1 band was confirmed following downregulation of RIPK1 with specific lentiviral <t>shRNA</t> in mouse PMN cultures (mPMNs). D , Quantification of RIPK1 protein levels. RIPK1 protein is significantly increased in Tg SOD1 G93A samples compared to Tg SOD1 WT ( p = 0.0279) and NTg ( p = 0.0033) mice. Results are presented as mean ± SEM. Statistical analysis was performed via one-way ANOVA followed by Tukey’s post hoc analysis; n = 3 biological replicates per genotype. E , Western blotting (RIPA) for RIPK3 showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Non-specific band at 47 kDa is designated as an asterisk (*). NTg spleen: positive control tissue, Ripk3 −/− spleen and Tg SOD1 G93A ; Ripk3 −/− spinal cord: negative control tissue. F , Western blotting (urea) for RIPK3 antibody showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Ripk3 −/− spinal cord: negative control tissue. G , Western blotting (RIPA) for MLKL showed no specific signal at the expected 55 kDa in spinal cord. NIH 3T3: positive control cell lysate. H , Western blotting (urea) for MLKL showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). NIH 3T3: positive control cell lysate. I , Western blotting for p-MLKL (RIPA) showed no signal at the expected 55 kDa. TSZ-treated L929 cells: control cell lysate.
    Shrna Containing Lentiviral Particles, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher lentiviral packaging
    <t>Lentiviral</t> Aβ 1–42 alters CX3CL1 levels and induces p-Tau
    Lentiviral Packaging, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 381 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    OriGene lentiviral packaging kit
    <t>Lentiviral</t> Aβ 1–42 alters CX3CL1 levels and induces p-Tau
    Lentiviral Packaging Kit, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Biowit Technologies lentiviral packaging kit
    <t>Lentiviral</t> Aβ 1–42 alters CX3CL1 levels and induces p-Tau
    Lentiviral Packaging Kit, supplied by Biowit Technologies, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    GenePharma Company lentiviral packaging kit
    <t>Lentiviral</t> Aβ 1–42 alters CX3CL1 levels and induces p-Tau
    Lentiviral Packaging Kit, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Shanghai Genechem lentiviral packaging kit
    Inhibitory effect of miR-223 on IGF1R/PI3K/Akt signaling pathway in vitro . (A) The GV259-miR-223 <t>lentiviral</t> vector was packaged and delivered to PC9/ER cells. Transduction efficiency was evaluated based on the fluorescent GFP signal. Fluorescent GFP signal was stably observed 72 h after infection. EV for control vector-infected cells (right panel) and miR-223 for GV259-miR-223-infected cells (left panel) are shown. Magnification, ×100. (B) miR-223 was overexpressed in PC9/ER cells as confirmed by real-time RT-PCR analysis. (C) Downregulation of IGF1R expression in PC9/ER-miR-223 cells was confirmed by real-time RT-PCR analysis. (D) IGF1R/Akt/p70S6K pathway was suppressed in the miR-223-overexpressing cells. Phosphorylation of Akt and p70S6K was significantly inhibited, according to western blot analysis. (E) miR-223 mediated inhibition of IGF1R/PI3K/Akt was reversed by agonist (IGF1) of IGF1R in miR-223 transfected cells. p-IGF1R and p-Atk were increased as shown in western blots. (F) The cytotoxicity and IC 50 values in PC9, PC9/ER, PC9/ER-EV, and PC9/ER-miR-223 cells. The cells were treated with the indicated concentrations of erlotinib for 72 h in medium containing 1% FBS. Cell viability and IC 50 values were determined using the CCK8 assay.
    Lentiviral Packaging Kit, supplied by Shanghai Genechem, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher lentiviral packaging kit
    MGAT5 restoration counteracts the suppressive effects of miR-124 on BC cell proliferation and migration. MDA-MB-231 and MCF-7 cells were transfected with miR-124 inhibitor, sh-MGAT5, sh-NC, and miR-124 inhibitor+sh-MGAT5 <t>lentiviral</t> vectors respectively to culture stably cell lines which followed by puromycin selection. A: Cell viability was measured using CCK-8 assays. B: EDU staining was performed and the percentage of EDU-positive cells were counted. Scale bar 100 µm. C: Representative photographs of the colony formation of MDA-MB-231 and MCF-7 cells, and related quantitative analysis. D: Cell migrations were determined using transwell. Scale bars, 50 µm. E: Cell migrations were determined using wound healing assay and the percentage of migrated was calculated. All data are shown as means ± SD of three separate experiments. * P
    Lentiviral Packaging Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Chronic GCN5L1 KD attenuated mitochondrial mass and protein levels via autophagic degradation. (A) Representative immunoblot of mitochondrial protein acetylation and the accumulation of autophagy mediators in isolated mitochondria following chronic lentiviral

    Journal: Journal of Cell Science

    Article Title: Restricted mitochondrial protein acetylation initiates mitochondrial autophagy

    doi: 10.1242/jcs.131300

    Figure Lengend Snippet: Chronic GCN5L1 KD attenuated mitochondrial mass and protein levels via autophagic degradation. (A) Representative immunoblot of mitochondrial protein acetylation and the accumulation of autophagy mediators in isolated mitochondria following chronic lentiviral

    Article Snippet: Lentiviral shRNA were generated in HEK293T cells using lentiviral Packaging Mix (Sigma).

    Techniques: Isolation

    Effects of shRNA on FMDV-IRES activity. a The shRNA-expressing lentiviral vector targeting the conserved region of FMDV-IRES (Lenti-FMDV-sh) or control vector (pLL3.7 alone) was used to infect HEK293 cells at various MOIs. After 14 days, firefly (FMDV-IRES activity) and Renilla (cap-dependent translation) luciferase activities were measured. To evaluate IRES-mediated translational activity, the ratio of IRES-mediated translation to cap-dependent translation was calculated. Experiments were performed in triplicate, and error bars indicate standard deviations. Multiple t tests were performed to calculate p values between cells with and without shRNA (** p

    Journal: Virus Genes

    Article Title: Silencing of the foot-and-mouth disease virus internal ribosomal entry site by targeting relatively conserved region among serotypes

    doi: 10.1007/s11262-019-01696-6

    Figure Lengend Snippet: Effects of shRNA on FMDV-IRES activity. a The shRNA-expressing lentiviral vector targeting the conserved region of FMDV-IRES (Lenti-FMDV-sh) or control vector (pLL3.7 alone) was used to infect HEK293 cells at various MOIs. After 14 days, firefly (FMDV-IRES activity) and Renilla (cap-dependent translation) luciferase activities were measured. To evaluate IRES-mediated translational activity, the ratio of IRES-mediated translation to cap-dependent translation was calculated. Experiments were performed in triplicate, and error bars indicate standard deviations. Multiple t tests were performed to calculate p values between cells with and without shRNA (** p

    Article Snippet: Lentiviral packaging was performed using MISSION Lentiviral Packaging Mix (Sigma-Aldrich, St. Louis, MO, USA), and infection with lentivirus was performed according to the manufacturer’s instructions.

    Techniques: shRNA, Activity Assay, Expressing, Plasmid Preparation, Luciferase

    SynNotch CD19-inducible ROR1 CAR-T cells are unable to rescue toxicity to ROR1 + normal tissues in the presence of circulating and bone marrow-resident CD19 + ROR1 + Raji tumors. ( A) Map of lentiviral constructs encoding CD19-Gal4VP64 synNotch Receptor, UAS inducible R11 ROR1 CAR, and constitutive R11 ROR1 CAR. (B) Raji tumor bioluminescence over time in NSG mice irradiated 250 R and treated with the indicated primary human T cells. n=6–7 per group. Two-way ANOVA with Tukey post-test (UT vs. ROR1 CAR-T: Day 13, 20, p

    Journal: Cancer cell

    Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

    doi: 10.1016/j.ccell.2019.02.003

    Figure Lengend Snippet: SynNotch CD19-inducible ROR1 CAR-T cells are unable to rescue toxicity to ROR1 + normal tissues in the presence of circulating and bone marrow-resident CD19 + ROR1 + Raji tumors. ( A) Map of lentiviral constructs encoding CD19-Gal4VP64 synNotch Receptor, UAS inducible R11 ROR1 CAR, and constitutive R11 ROR1 CAR. (B) Raji tumor bioluminescence over time in NSG mice irradiated 250 R and treated with the indicated primary human T cells. n=6–7 per group. Two-way ANOVA with Tukey post-test (UT vs. ROR1 CAR-T: Day 13, 20, p

    Article Snippet: Lenti-X cells for lentiviral packaging were purchased from Clontech.

    Techniques: Construct, Mouse Assay, Irradiation

    Intein splicing kinetics. ( A ) Autoradiogram after SDS/PAGE and anti-HA immunoprecipitation of lysates from GFP-RecA-HY lentivirus-infected E36 cells pretreated with epoxomicin (5 μM, 2 h) subjected to [ 35 S]-Met/Cys pulse label followed by a “cold”

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Using intein catalysis to probe the origin of major histocompatibility complex class I-presented peptides

    doi: 10.1073/pnas.1210271109

    Figure Lengend Snippet: Intein splicing kinetics. ( A ) Autoradiogram after SDS/PAGE and anti-HA immunoprecipitation of lysates from GFP-RecA-HY lentivirus-infected E36 cells pretreated with epoxomicin (5 μM, 2 h) subjected to [ 35 S]-Met/Cys pulse label followed by a “cold”

    Article Snippet: Vesicular stomatitis virus G protein (VSV-G) pseudotyped lentivirus particles were generated by cotransfecting the lentiviral vector with the Open Biosystems lentivirus packaging mix into LentiX 293T cells (Clontech).

    Techniques: SDS Page, Immunoprecipitation, Infection

    Upregulation of core necroptosis components in the spinal cord of symptomatic Tg SOD1 G93A mice. Lumbar spinal cords, from 12- and 15-week-old Tg SOD1 G93A , WT SOD1, and NTg mice, were isolated and processed for mRNA and protein (RIPA or urea extraction) expression of RIPK1, RIPK3, MLKL, and p-MLKL. A , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA from 12-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk3 in Tg SOD1 G93A compared to Tg SOD1 WT ( p = 0.0021) and NTg ( p = 0.0076) at 12 weeks. B , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA in spinal cords of 15-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk1 ( p = 0.0009, vs Tg SOD1 WT ; p = 0.0020, vs NTg) and Ripk3 ( p = 0.0115; vs Tg SOD1 WT , p = 0.0067; vs NTg) but not for Mlkl in Tg SOD1 G93A compared to Tg SOD1 WT and NTg mice at 15 weeks. C , Western blotting (RIPA) for RIPK1 in spinal cord of NTg, Tg SOD1 WT , and Tg SOD1 G93A 15-week-old mice. β-ACTIN, GAPDH: loading control. Specificity of the RIPK1 band was confirmed following downregulation of RIPK1 with specific lentiviral shRNA in mouse PMN cultures (mPMNs). D , Quantification of RIPK1 protein levels. RIPK1 protein is significantly increased in Tg SOD1 G93A samples compared to Tg SOD1 WT ( p = 0.0279) and NTg ( p = 0.0033) mice. Results are presented as mean ± SEM. Statistical analysis was performed via one-way ANOVA followed by Tukey’s post hoc analysis; n = 3 biological replicates per genotype. E , Western blotting (RIPA) for RIPK3 showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Non-specific band at 47 kDa is designated as an asterisk (*). NTg spleen: positive control tissue, Ripk3 −/− spleen and Tg SOD1 G93A ; Ripk3 −/− spinal cord: negative control tissue. F , Western blotting (urea) for RIPK3 antibody showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Ripk3 −/− spinal cord: negative control tissue. G , Western blotting (RIPA) for MLKL showed no specific signal at the expected 55 kDa in spinal cord. NIH 3T3: positive control cell lysate. H , Western blotting (urea) for MLKL showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). NIH 3T3: positive control cell lysate. I , Western blotting for p-MLKL (RIPA) showed no signal at the expected 55 kDa. TSZ-treated L929 cells: control cell lysate.

    Journal: eNeuro

    Article Title: Deletion of Ripk3 Prevents Motor Neuron Death In Vitro but not In Vivo

    doi: 10.1523/ENEURO.0308-18.2018

    Figure Lengend Snippet: Upregulation of core necroptosis components in the spinal cord of symptomatic Tg SOD1 G93A mice. Lumbar spinal cords, from 12- and 15-week-old Tg SOD1 G93A , WT SOD1, and NTg mice, were isolated and processed for mRNA and protein (RIPA or urea extraction) expression of RIPK1, RIPK3, MLKL, and p-MLKL. A , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA from 12-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk3 in Tg SOD1 G93A compared to Tg SOD1 WT ( p = 0.0021) and NTg ( p = 0.0076) at 12 weeks. B , Quantification of Ripk1 , Ripk3 , and Mlkl mRNA in spinal cords of 15-week-old mice . Gapdh : housekeeping gene. A significant increase was detected for Ripk1 ( p = 0.0009, vs Tg SOD1 WT ; p = 0.0020, vs NTg) and Ripk3 ( p = 0.0115; vs Tg SOD1 WT , p = 0.0067; vs NTg) but not for Mlkl in Tg SOD1 G93A compared to Tg SOD1 WT and NTg mice at 15 weeks. C , Western blotting (RIPA) for RIPK1 in spinal cord of NTg, Tg SOD1 WT , and Tg SOD1 G93A 15-week-old mice. β-ACTIN, GAPDH: loading control. Specificity of the RIPK1 band was confirmed following downregulation of RIPK1 with specific lentiviral shRNA in mouse PMN cultures (mPMNs). D , Quantification of RIPK1 protein levels. RIPK1 protein is significantly increased in Tg SOD1 G93A samples compared to Tg SOD1 WT ( p = 0.0279) and NTg ( p = 0.0033) mice. Results are presented as mean ± SEM. Statistical analysis was performed via one-way ANOVA followed by Tukey’s post hoc analysis; n = 3 biological replicates per genotype. E , Western blotting (RIPA) for RIPK3 showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Non-specific band at 47 kDa is designated as an asterisk (*). NTg spleen: positive control tissue, Ripk3 −/− spleen and Tg SOD1 G93A ; Ripk3 −/− spinal cord: negative control tissue. F , Western blotting (urea) for RIPK3 antibody showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). Ripk3 −/− spinal cord: negative control tissue. G , Western blotting (RIPA) for MLKL showed no specific signal at the expected 55 kDa in spinal cord. NIH 3T3: positive control cell lysate. H , Western blotting (urea) for MLKL showed no specific signal at the expected 55 kDa in spinal cord (NTg and Tg SOD1 G93A ). NIH 3T3: positive control cell lysate. I , Western blotting for p-MLKL (RIPA) showed no signal at the expected 55 kDa. TSZ-treated L929 cells: control cell lysate.

    Article Snippet: For mixed-lineage kinase domain-like (Mlkl) silencing, L929 cells were infected with lentivirus-containing shRNA against Mlkl (360819; Sigma Mission) at a multiplicity of infection (MOI) 100.

    Techniques: Mouse Assay, Isolation, Expressing, Western Blot, shRNA, Positive Control, Negative Control

    Lentiviral Aβ 1–42 alters CX3CL1 levels and induces p-Tau

    Journal: Experimental neurology

    Article Title: Fractalkine signaling and Tau hyper-phosphorylation are associated with autophagic alterations in lentiviral Tau and Aβ1–42 gene transfer models

    doi: 10.1016/j.expneurol.2013.01.009

    Figure Lengend Snippet: Lentiviral Aβ 1–42 alters CX3CL1 levels and induces p-Tau

    Article Snippet: Lentiviral packaging was performed according to manufacturer’s protocol (Invitrogen, Inc).

    Techniques:

    Microglia are activated 2 weeks post-injection with lentiviral Aβ 1–42

    Journal: Experimental neurology

    Article Title: Fractalkine signaling and Tau hyper-phosphorylation are associated with autophagic alterations in lentiviral Tau and Aβ1–42 gene transfer models

    doi: 10.1016/j.expneurol.2013.01.009

    Figure Lengend Snippet: Microglia are activated 2 weeks post-injection with lentiviral Aβ 1–42

    Article Snippet: Lentiviral packaging was performed according to manufacturer’s protocol (Invitrogen, Inc).

    Techniques: Injection

    Inhibitory effect of miR-223 on IGF1R/PI3K/Akt signaling pathway in vitro . (A) The GV259-miR-223 lentiviral vector was packaged and delivered to PC9/ER cells. Transduction efficiency was evaluated based on the fluorescent GFP signal. Fluorescent GFP signal was stably observed 72 h after infection. EV for control vector-infected cells (right panel) and miR-223 for GV259-miR-223-infected cells (left panel) are shown. Magnification, ×100. (B) miR-223 was overexpressed in PC9/ER cells as confirmed by real-time RT-PCR analysis. (C) Downregulation of IGF1R expression in PC9/ER-miR-223 cells was confirmed by real-time RT-PCR analysis. (D) IGF1R/Akt/p70S6K pathway was suppressed in the miR-223-overexpressing cells. Phosphorylation of Akt and p70S6K was significantly inhibited, according to western blot analysis. (E) miR-223 mediated inhibition of IGF1R/PI3K/Akt was reversed by agonist (IGF1) of IGF1R in miR-223 transfected cells. p-IGF1R and p-Atk were increased as shown in western blots. (F) The cytotoxicity and IC 50 values in PC9, PC9/ER, PC9/ER-EV, and PC9/ER-miR-223 cells. The cells were treated with the indicated concentrations of erlotinib for 72 h in medium containing 1% FBS. Cell viability and IC 50 values were determined using the CCK8 assay.

    Journal: International Journal of Oncology

    Article Title: miR-223 reverses the resistance of EGFR-TKIs through IGF1R/PI3K/Akt signaling pathway

    doi: 10.3892/ijo.2016.3401

    Figure Lengend Snippet: Inhibitory effect of miR-223 on IGF1R/PI3K/Akt signaling pathway in vitro . (A) The GV259-miR-223 lentiviral vector was packaged and delivered to PC9/ER cells. Transduction efficiency was evaluated based on the fluorescent GFP signal. Fluorescent GFP signal was stably observed 72 h after infection. EV for control vector-infected cells (right panel) and miR-223 for GV259-miR-223-infected cells (left panel) are shown. Magnification, ×100. (B) miR-223 was overexpressed in PC9/ER cells as confirmed by real-time RT-PCR analysis. (C) Downregulation of IGF1R expression in PC9/ER-miR-223 cells was confirmed by real-time RT-PCR analysis. (D) IGF1R/Akt/p70S6K pathway was suppressed in the miR-223-overexpressing cells. Phosphorylation of Akt and p70S6K was significantly inhibited, according to western blot analysis. (E) miR-223 mediated inhibition of IGF1R/PI3K/Akt was reversed by agonist (IGF1) of IGF1R in miR-223 transfected cells. p-IGF1R and p-Atk were increased as shown in western blots. (F) The cytotoxicity and IC 50 values in PC9, PC9/ER, PC9/ER-EV, and PC9/ER-miR-223 cells. The cells were treated with the indicated concentrations of erlotinib for 72 h in medium containing 1% FBS. Cell viability and IC 50 values were determined using the CCK8 assay.

    Article Snippet: Construction of stable cell lines with overexpressed miR-223 To stably upregulate miR-223 expression in PC9/ER cells or PC9/CD133+ , lentivirus carrying the miR-223 or negative control vector (empty viral vector; EV) was packaged using the lentiviral packaging kit (Shanghai Genechem Co., Ltd., Shanghai, China), according to the manufacturer's instructions.

    Techniques: In Vitro, Plasmid Preparation, Transduction, Stable Transfection, Infection, Quantitative RT-PCR, Expressing, Western Blot, Inhibition, Transfection, CCK-8 Assay

    miR-223 inhibits the IGF1R/PI3K/Akt signaling pathway in CD133 + subpopulation cells. (A) The GV259-miR-223 lentiviral vector was packaged and delivered to PC9/CD133 + cells. Transduction efficiency was evaluated based on fluorescent GFP signal. Fluorescent GFP signal was stably observed 72 h after infection. EV for control vector-infected cells (a) and miR-223 for GV259-miR-223-infected cells (b). Magnification, ×100. (B) Transduction efficiency was evaluated by real-time RT-PCR analysis, and expression of miR-223 in PC9/CD133 + -miR-223 cells was 12.5-fold that in PC9/CD133 + cells. (C) Cytotoxicity and IC 50 values of erlotinib in PC9,PC9/CD133 + , PC9/CD133 + -miR-223, and PC9/CD133 + -EV cells. Cells were treated with indicated concentrations of erlotinib for 72 h in medium containing 1% FBS. Cell viability and IC 50 values in the treated cells were determined using the CCK8 assay. (D) Western blot analysis of the key molecules of IGF1R/PI3K/Akt signaling pathway in the erlotinib-treated (0.01 μM for 72 h) PC9, PC9/CD133 + , PC9/CD133 + -miR-223, and PC9/CD133 + -EV cells.

    Journal: International Journal of Oncology

    Article Title: miR-223 reverses the resistance of EGFR-TKIs through IGF1R/PI3K/Akt signaling pathway

    doi: 10.3892/ijo.2016.3401

    Figure Lengend Snippet: miR-223 inhibits the IGF1R/PI3K/Akt signaling pathway in CD133 + subpopulation cells. (A) The GV259-miR-223 lentiviral vector was packaged and delivered to PC9/CD133 + cells. Transduction efficiency was evaluated based on fluorescent GFP signal. Fluorescent GFP signal was stably observed 72 h after infection. EV for control vector-infected cells (a) and miR-223 for GV259-miR-223-infected cells (b). Magnification, ×100. (B) Transduction efficiency was evaluated by real-time RT-PCR analysis, and expression of miR-223 in PC9/CD133 + -miR-223 cells was 12.5-fold that in PC9/CD133 + cells. (C) Cytotoxicity and IC 50 values of erlotinib in PC9,PC9/CD133 + , PC9/CD133 + -miR-223, and PC9/CD133 + -EV cells. Cells were treated with indicated concentrations of erlotinib for 72 h in medium containing 1% FBS. Cell viability and IC 50 values in the treated cells were determined using the CCK8 assay. (D) Western blot analysis of the key molecules of IGF1R/PI3K/Akt signaling pathway in the erlotinib-treated (0.01 μM for 72 h) PC9, PC9/CD133 + , PC9/CD133 + -miR-223, and PC9/CD133 + -EV cells.

    Article Snippet: Construction of stable cell lines with overexpressed miR-223 To stably upregulate miR-223 expression in PC9/ER cells or PC9/CD133+ , lentivirus carrying the miR-223 or negative control vector (empty viral vector; EV) was packaged using the lentiviral packaging kit (Shanghai Genechem Co., Ltd., Shanghai, China), according to the manufacturer's instructions.

    Techniques: Plasmid Preparation, Transduction, Stable Transfection, Infection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Western Blot

    MGAT5 restoration counteracts the suppressive effects of miR-124 on BC cell proliferation and migration. MDA-MB-231 and MCF-7 cells were transfected with miR-124 inhibitor, sh-MGAT5, sh-NC, and miR-124 inhibitor+sh-MGAT5 lentiviral vectors respectively to culture stably cell lines which followed by puromycin selection. A: Cell viability was measured using CCK-8 assays. B: EDU staining was performed and the percentage of EDU-positive cells were counted. Scale bar 100 µm. C: Representative photographs of the colony formation of MDA-MB-231 and MCF-7 cells, and related quantitative analysis. D: Cell migrations were determined using transwell. Scale bars, 50 µm. E: Cell migrations were determined using wound healing assay and the percentage of migrated was calculated. All data are shown as means ± SD of three separate experiments. * P

    Journal: American Journal of Cancer Research

    Article Title: Decreased miR-124-3p promoted breast cancer proliferation and metastasis by targeting MGAT5

    doi:

    Figure Lengend Snippet: MGAT5 restoration counteracts the suppressive effects of miR-124 on BC cell proliferation and migration. MDA-MB-231 and MCF-7 cells were transfected with miR-124 inhibitor, sh-MGAT5, sh-NC, and miR-124 inhibitor+sh-MGAT5 lentiviral vectors respectively to culture stably cell lines which followed by puromycin selection. A: Cell viability was measured using CCK-8 assays. B: EDU staining was performed and the percentage of EDU-positive cells were counted. Scale bar 100 µm. C: Representative photographs of the colony formation of MDA-MB-231 and MCF-7 cells, and related quantitative analysis. D: Cell migrations were determined using transwell. Scale bars, 50 µm. E: Cell migrations were determined using wound healing assay and the percentage of migrated was calculated. All data are shown as means ± SD of three separate experiments. * P

    Article Snippet: The lentiviral packaging kit was purchased from Open Biosystems (Genomeditech).

    Techniques: Migration, Multiple Displacement Amplification, Transfection, Stable Transfection, Selection, CCK-8 Assay, Staining, Wound Healing Assay

    MGAT5 restoration counteracts the suppressive effects of miR-124 on BC cell proliferation and migration. MDA-MB-231 and MCF-7 cells were transfected with miR-124, MGAT5, NC, and miR-124+MGAT5 lentiviral vectors respectively to culture stably cell lines which followed by puromycin selection. A: Cell viability was measured using CCK-8 assays. B: EDU staining was performed and the percentage of EDU-positive cells were counted. Scale bar 100 µm. C: Representative photographs of the colony formation of MDA-MB-231 and MCF-7 cells, and related quantitative analysis. D: Cell migrations were determined using transwell. Scale bars, 50 µm. E: Cell migrations were determined using wound healing assay and the percentage of migrated was calculated. All data are shown as means ± SD of three separate experiments. * P

    Journal: American Journal of Cancer Research

    Article Title: Decreased miR-124-3p promoted breast cancer proliferation and metastasis by targeting MGAT5

    doi:

    Figure Lengend Snippet: MGAT5 restoration counteracts the suppressive effects of miR-124 on BC cell proliferation and migration. MDA-MB-231 and MCF-7 cells were transfected with miR-124, MGAT5, NC, and miR-124+MGAT5 lentiviral vectors respectively to culture stably cell lines which followed by puromycin selection. A: Cell viability was measured using CCK-8 assays. B: EDU staining was performed and the percentage of EDU-positive cells were counted. Scale bar 100 µm. C: Representative photographs of the colony formation of MDA-MB-231 and MCF-7 cells, and related quantitative analysis. D: Cell migrations were determined using transwell. Scale bars, 50 µm. E: Cell migrations were determined using wound healing assay and the percentage of migrated was calculated. All data are shown as means ± SD of three separate experiments. * P

    Article Snippet: The lentiviral packaging kit was purchased from Open Biosystems (Genomeditech).

    Techniques: Migration, Multiple Displacement Amplification, Transfection, Stable Transfection, Selection, CCK-8 Assay, Staining, Wound Healing Assay