anti tau  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau
    Anti Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti tau  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau
    Anti Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti tau  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau
    Anti Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti lrp1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti lrp1
    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
    Anti Lrp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons"

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0113237

    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying LRP1-shRNA or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
    Figure Legend Snippet: Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying LRP1-shRNA or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Techniques Used: Cell Culture, Infection, shRNA, In Vitro, Expressing, Western Blot, MTT Assay, Real-time Polymerase Chain Reaction

    ( A ) Brain lysates from wild-type mice were immune-precipitated using specific antibodies against LRP1, GluA1, GluA2/3 or PSD95, and their interactions were examined by Western blot ( B–E ). After infection with control NT-shRNA or LRP1-shRNA, control and LRP1-knockdown neurons were treated with cycloheximide (CHX), and the levels of GluA1 ( C ), GluA2/3 ( D ) and PSD95 ( E ) were analyzed by Western blot at different time points. ( F ) LRP1-knockdown neurons were treated with DMSO (control), proteasomal inhibitor lactacystin (Lac; 10 µM) or lysosomal inhibitor bafilomycin A1 (BA1; 5 nM) in addition to CHX. ( G ) GluA1 and PSD95 levels were analyzed by Western blot, and densitometrically quantified. The data are plotted as mean ± SD (n = 3). *, p<0.05; **, p<0.01.
    Figure Legend Snippet: ( A ) Brain lysates from wild-type mice were immune-precipitated using specific antibodies against LRP1, GluA1, GluA2/3 or PSD95, and their interactions were examined by Western blot ( B–E ). After infection with control NT-shRNA or LRP1-shRNA, control and LRP1-knockdown neurons were treated with cycloheximide (CHX), and the levels of GluA1 ( C ), GluA2/3 ( D ) and PSD95 ( E ) were analyzed by Western blot at different time points. ( F ) LRP1-knockdown neurons were treated with DMSO (control), proteasomal inhibitor lactacystin (Lac; 10 µM) or lysosomal inhibitor bafilomycin A1 (BA1; 5 nM) in addition to CHX. ( G ) GluA1 and PSD95 levels were analyzed by Western blot, and densitometrically quantified. The data are plotted as mean ± SD (n = 3). *, p<0.05; **, p<0.01.

    Techniques Used: Western Blot, Infection, shRNA

    Primary mouse cortical neurons were infected with lentivirus carrying LRP1-shRNA or NT-shRNA for 4 days. Cell surface proteins were labeled with biotin in live neurons, and the cell lysates were precipitated with streptavidin beads. ( A, B ) The precipitates and total cell lysates were examined by Western blot to detect cell surface GluA1 and total GluA1, respectively. The ratio of surface GluA1 versus total GluA1 was quantified ( A ). Similarly, ratio of surface GluA2/3 versus total GluA2/3 was analyzed ( B ). ( C ) In control and LRP1-knockdown neurons, the expression of total GluA1 and phosphorylated GluA1 (pSer-845 and pSer-831) were analyzed by Western blot. The phosphorylation at Ser-845 ( D ) and Ser-831( E ) sites of GluA1 versus total GluA1 were quantified. The data are plotted as mean ± SD (n = 3). N.S., not significant; *, p<0.05; **, p<0.01.
    Figure Legend Snippet: Primary mouse cortical neurons were infected with lentivirus carrying LRP1-shRNA or NT-shRNA for 4 days. Cell surface proteins were labeled with biotin in live neurons, and the cell lysates were precipitated with streptavidin beads. ( A, B ) The precipitates and total cell lysates were examined by Western blot to detect cell surface GluA1 and total GluA1, respectively. The ratio of surface GluA1 versus total GluA1 was quantified ( A ). Similarly, ratio of surface GluA2/3 versus total GluA2/3 was analyzed ( B ). ( C ) In control and LRP1-knockdown neurons, the expression of total GluA1 and phosphorylated GluA1 (pSer-845 and pSer-831) were analyzed by Western blot. The phosphorylation at Ser-845 ( D ) and Ser-831( E ) sites of GluA1 versus total GluA1 were quantified. The data are plotted as mean ± SD (n = 3). N.S., not significant; *, p<0.05; **, p<0.01.

    Techniques Used: Infection, shRNA, Labeling, Western Blot, Expressing

    Primary mouse neurons were first infected with lentivirus carrying control vector or GluA1 plasmid, and then with lentivirus carrying NT-shRNA or LRP1-shRNA ( A ). Expression levels of LRP1 ( B ) and GluA1 ( C ) were detected by Western blot. ( D ) Calcium influx detected with the fluorescence microplate reader using Fluo-4 AM as a fluorescent indicator of intracellular calcium concentration in neurons after stimulation of AMPA in the presence of NMDAR antagonist. The scale bar represents 200 µm. ( E ) Calcium fluorescence intensities were measured with the excitation and emission wavelengths set at 494 and 535 nm, respectively. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
    Figure Legend Snippet: Primary mouse neurons were first infected with lentivirus carrying control vector or GluA1 plasmid, and then with lentivirus carrying NT-shRNA or LRP1-shRNA ( A ). Expression levels of LRP1 ( B ) and GluA1 ( C ) were detected by Western blot. ( D ) Calcium influx detected with the fluorescence microplate reader using Fluo-4 AM as a fluorescent indicator of intracellular calcium concentration in neurons after stimulation of AMPA in the presence of NMDAR antagonist. The scale bar represents 200 µm. ( E ) Calcium fluorescence intensities were measured with the excitation and emission wavelengths set at 494 and 535 nm, respectively. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Techniques Used: Infection, Plasmid Preparation, shRNA, Expressing, Western Blot, Fluorescence, Concentration Assay

    Mouse primary neurons were infected with lentivirus carrying control vector or GluA1 cDNA and lentivirus carrying NT-shRNA or LRP1-shRNA. Control and LRP1-suppressed neurons with or without forced GluA1 expression were stained with anti-MAP2 antibody and their neurite outgrowth ( A ; scale bar = 25 µm) and filopodia formation ( B ; scale bar = 15 µm) were observed using confocal microscopy. Total outgrowth ( C ), mean process length ( D ) and Filopodia density ( E ) were quantified by MetaMorph software. The data are plotted as mean ± SEM. N.S., Not significant; *, p<0.05; **, p<0.01.
    Figure Legend Snippet: Mouse primary neurons were infected with lentivirus carrying control vector or GluA1 cDNA and lentivirus carrying NT-shRNA or LRP1-shRNA. Control and LRP1-suppressed neurons with or without forced GluA1 expression were stained with anti-MAP2 antibody and their neurite outgrowth ( A ; scale bar = 25 µm) and filopodia formation ( B ; scale bar = 15 µm) were observed using confocal microscopy. Total outgrowth ( C ), mean process length ( D ) and Filopodia density ( E ) were quantified by MetaMorph software. The data are plotted as mean ± SEM. N.S., Not significant; *, p<0.05; **, p<0.01.

    Techniques Used: Infection, Plasmid Preparation, shRNA, Expressing, Staining, Confocal Microscopy, Software

    p tau  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p tau
    P Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti tau ps404 d2z4g  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau ps404 d2z4g
    Anti Tau Ps404 D2z4g, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti tau c terminus d1m9x  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau c terminus d1m9x
    Anti Tau C Terminus D1m9x, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti tau pt205 e7d3e  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti tau pt205 e7d3e
    Anti Tau Pt205 E7d3e, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tau ps262  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc tau ps262
    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, <t>Tau-pS262,</t> Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.
    Tau Ps262, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets"

    Article Title: Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.204336

    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.
    Figure Legend Snippet: 2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Techniques Used: De-Phosphorylation Assay, Activity Assay, Western Blot, Inhibition, Molecular Weight

    tau ps396  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc tau ps396
    2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and <t>tau-pS396</t> in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.
    Tau Ps396, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets"

    Article Title: Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.204336

    2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and tau-pS396 in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.
    Figure Legend Snippet: 2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and tau-pS396 in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.

    Techniques Used: Transfection, Plasmid Preparation, Western Blot, Molecular Weight

    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.
    Figure Legend Snippet: 2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Techniques Used: De-Phosphorylation Assay, Activity Assay, Western Blot, Inhibition, Molecular Weight

    tau  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc tau
    Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti tau
    Anti Tau, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti lrp1
    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
    Anti Lrp1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc p tau
    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
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    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
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    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
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    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying <t>LRP1-shRNA</t> or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.
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    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, <t>Tau-pS262,</t> Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.
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    2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and <t>tau-pS396</t> in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.
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    2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and <t>tau-pS396</t> in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.
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    Image Search Results


    Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying LRP1-shRNA or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Journal: PLoS ONE

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    doi: 10.1371/journal.pone.0113237

    Figure Lengend Snippet: Primary cortical neurons cultured from C57Bl/6 mice were infected with lentivirus carrying LRP1-shRNA or control NT-shRNA on day 8 in vitro (DIV) and then harvested after 2 or 4 days of infection. The expression level of LRP1 in neurons was detected by Western blot ( A ), and densitometrically quantified ( B ). ( C ) The cell viability of neurons was assessed by MTT assay at 2 or 4 days following infection. In LRP1-knockdown neurons, the expression levels of PSD95 ( D , E ), GluA1 ( D , F ), and GluA2/3 ( D , G ) at 4 days post-infection were detected by Western blot and densitometrically quantified. In addition, the mRNA levels of PSD95 ( H ) and GluA1 ( I ) were also analyzed by quantitative real-time PCR. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Article Snippet: The primary antibodies used were: anti-GluA1 (1∶300, Millipore), anti-GluA2/3 (1∶500, Millipore), anti-pS831-GluA1 (1∶500, Epitomics, Burlingame, CA), anti-pS845-GluA1 (1∶500, Epitomics), anti-LRP1 (1∶2000, made in house) , anti-PSD95 (1∶1000, Cell Signaling, Danvers, MA) and anti-β actin (1∶10, 000, Sigma-Aldrich).

    Techniques: Cell Culture, Infection, shRNA, In Vitro, Expressing, Western Blot, MTT Assay, Real-time Polymerase Chain Reaction

    ( A ) Brain lysates from wild-type mice were immune-precipitated using specific antibodies against LRP1, GluA1, GluA2/3 or PSD95, and their interactions were examined by Western blot ( B–E ). After infection with control NT-shRNA or LRP1-shRNA, control and LRP1-knockdown neurons were treated with cycloheximide (CHX), and the levels of GluA1 ( C ), GluA2/3 ( D ) and PSD95 ( E ) were analyzed by Western blot at different time points. ( F ) LRP1-knockdown neurons were treated with DMSO (control), proteasomal inhibitor lactacystin (Lac; 10 µM) or lysosomal inhibitor bafilomycin A1 (BA1; 5 nM) in addition to CHX. ( G ) GluA1 and PSD95 levels were analyzed by Western blot, and densitometrically quantified. The data are plotted as mean ± SD (n = 3). *, p<0.05; **, p<0.01.

    Journal: PLoS ONE

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    doi: 10.1371/journal.pone.0113237

    Figure Lengend Snippet: ( A ) Brain lysates from wild-type mice were immune-precipitated using specific antibodies against LRP1, GluA1, GluA2/3 or PSD95, and their interactions were examined by Western blot ( B–E ). After infection with control NT-shRNA or LRP1-shRNA, control and LRP1-knockdown neurons were treated with cycloheximide (CHX), and the levels of GluA1 ( C ), GluA2/3 ( D ) and PSD95 ( E ) were analyzed by Western blot at different time points. ( F ) LRP1-knockdown neurons were treated with DMSO (control), proteasomal inhibitor lactacystin (Lac; 10 µM) or lysosomal inhibitor bafilomycin A1 (BA1; 5 nM) in addition to CHX. ( G ) GluA1 and PSD95 levels were analyzed by Western blot, and densitometrically quantified. The data are plotted as mean ± SD (n = 3). *, p<0.05; **, p<0.01.

    Article Snippet: The primary antibodies used were: anti-GluA1 (1∶300, Millipore), anti-GluA2/3 (1∶500, Millipore), anti-pS831-GluA1 (1∶500, Epitomics, Burlingame, CA), anti-pS845-GluA1 (1∶500, Epitomics), anti-LRP1 (1∶2000, made in house) , anti-PSD95 (1∶1000, Cell Signaling, Danvers, MA) and anti-β actin (1∶10, 000, Sigma-Aldrich).

    Techniques: Western Blot, Infection, shRNA

    Primary mouse cortical neurons were infected with lentivirus carrying LRP1-shRNA or NT-shRNA for 4 days. Cell surface proteins were labeled with biotin in live neurons, and the cell lysates were precipitated with streptavidin beads. ( A, B ) The precipitates and total cell lysates were examined by Western blot to detect cell surface GluA1 and total GluA1, respectively. The ratio of surface GluA1 versus total GluA1 was quantified ( A ). Similarly, ratio of surface GluA2/3 versus total GluA2/3 was analyzed ( B ). ( C ) In control and LRP1-knockdown neurons, the expression of total GluA1 and phosphorylated GluA1 (pSer-845 and pSer-831) were analyzed by Western blot. The phosphorylation at Ser-845 ( D ) and Ser-831( E ) sites of GluA1 versus total GluA1 were quantified. The data are plotted as mean ± SD (n = 3). N.S., not significant; *, p<0.05; **, p<0.01.

    Journal: PLoS ONE

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    doi: 10.1371/journal.pone.0113237

    Figure Lengend Snippet: Primary mouse cortical neurons were infected with lentivirus carrying LRP1-shRNA or NT-shRNA for 4 days. Cell surface proteins were labeled with biotin in live neurons, and the cell lysates were precipitated with streptavidin beads. ( A, B ) The precipitates and total cell lysates were examined by Western blot to detect cell surface GluA1 and total GluA1, respectively. The ratio of surface GluA1 versus total GluA1 was quantified ( A ). Similarly, ratio of surface GluA2/3 versus total GluA2/3 was analyzed ( B ). ( C ) In control and LRP1-knockdown neurons, the expression of total GluA1 and phosphorylated GluA1 (pSer-845 and pSer-831) were analyzed by Western blot. The phosphorylation at Ser-845 ( D ) and Ser-831( E ) sites of GluA1 versus total GluA1 were quantified. The data are plotted as mean ± SD (n = 3). N.S., not significant; *, p<0.05; **, p<0.01.

    Article Snippet: The primary antibodies used were: anti-GluA1 (1∶300, Millipore), anti-GluA2/3 (1∶500, Millipore), anti-pS831-GluA1 (1∶500, Epitomics, Burlingame, CA), anti-pS845-GluA1 (1∶500, Epitomics), anti-LRP1 (1∶2000, made in house) , anti-PSD95 (1∶1000, Cell Signaling, Danvers, MA) and anti-β actin (1∶10, 000, Sigma-Aldrich).

    Techniques: Infection, shRNA, Labeling, Western Blot, Expressing

    Primary mouse neurons were first infected with lentivirus carrying control vector or GluA1 plasmid, and then with lentivirus carrying NT-shRNA or LRP1-shRNA ( A ). Expression levels of LRP1 ( B ) and GluA1 ( C ) were detected by Western blot. ( D ) Calcium influx detected with the fluorescence microplate reader using Fluo-4 AM as a fluorescent indicator of intracellular calcium concentration in neurons after stimulation of AMPA in the presence of NMDAR antagonist. The scale bar represents 200 µm. ( E ) Calcium fluorescence intensities were measured with the excitation and emission wavelengths set at 494 and 535 nm, respectively. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Journal: PLoS ONE

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    doi: 10.1371/journal.pone.0113237

    Figure Lengend Snippet: Primary mouse neurons were first infected with lentivirus carrying control vector or GluA1 plasmid, and then with lentivirus carrying NT-shRNA or LRP1-shRNA ( A ). Expression levels of LRP1 ( B ) and GluA1 ( C ) were detected by Western blot. ( D ) Calcium influx detected with the fluorescence microplate reader using Fluo-4 AM as a fluorescent indicator of intracellular calcium concentration in neurons after stimulation of AMPA in the presence of NMDAR antagonist. The scale bar represents 200 µm. ( E ) Calcium fluorescence intensities were measured with the excitation and emission wavelengths set at 494 and 535 nm, respectively. The data are plotted as mean ± SD (n = 3). N.S., Not significant; **, p<0.01.

    Article Snippet: The primary antibodies used were: anti-GluA1 (1∶300, Millipore), anti-GluA2/3 (1∶500, Millipore), anti-pS831-GluA1 (1∶500, Epitomics, Burlingame, CA), anti-pS845-GluA1 (1∶500, Epitomics), anti-LRP1 (1∶2000, made in house) , anti-PSD95 (1∶1000, Cell Signaling, Danvers, MA) and anti-β actin (1∶10, 000, Sigma-Aldrich).

    Techniques: Infection, Plasmid Preparation, shRNA, Expressing, Western Blot, Fluorescence, Concentration Assay

    Mouse primary neurons were infected with lentivirus carrying control vector or GluA1 cDNA and lentivirus carrying NT-shRNA or LRP1-shRNA. Control and LRP1-suppressed neurons with or without forced GluA1 expression were stained with anti-MAP2 antibody and their neurite outgrowth ( A ; scale bar = 25 µm) and filopodia formation ( B ; scale bar = 15 µm) were observed using confocal microscopy. Total outgrowth ( C ), mean process length ( D ) and Filopodia density ( E ) were quantified by MetaMorph software. The data are plotted as mean ± SEM. N.S., Not significant; *, p<0.05; **, p<0.01.

    Journal: PLoS ONE

    Article Title: Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Regulates the Stability and Function of GluA1 α-Amino-3-Hydroxy-5-Methyl-4-Isoxazole Propionic Acid (AMPA) Receptor in Neurons

    doi: 10.1371/journal.pone.0113237

    Figure Lengend Snippet: Mouse primary neurons were infected with lentivirus carrying control vector or GluA1 cDNA and lentivirus carrying NT-shRNA or LRP1-shRNA. Control and LRP1-suppressed neurons with or without forced GluA1 expression were stained with anti-MAP2 antibody and their neurite outgrowth ( A ; scale bar = 25 µm) and filopodia formation ( B ; scale bar = 15 µm) were observed using confocal microscopy. Total outgrowth ( C ), mean process length ( D ) and Filopodia density ( E ) were quantified by MetaMorph software. The data are plotted as mean ± SEM. N.S., Not significant; *, p<0.05; **, p<0.01.

    Article Snippet: The primary antibodies used were: anti-GluA1 (1∶300, Millipore), anti-GluA2/3 (1∶500, Millipore), anti-pS831-GluA1 (1∶500, Epitomics, Burlingame, CA), anti-pS845-GluA1 (1∶500, Epitomics), anti-LRP1 (1∶2000, made in house) , anti-PSD95 (1∶1000, Cell Signaling, Danvers, MA) and anti-β actin (1∶10, 000, Sigma-Aldrich).

    Techniques: Infection, Plasmid Preparation, shRNA, Expressing, Staining, Confocal Microscopy, Software

    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Journal: Aging (Albany NY)

    Article Title: Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets

    doi: 10.18632/aging.204336

    Figure Lengend Snippet: 2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Article Snippet: The primary antibodies utilized in this study are as follows: GSK-3β (21002, Signalway Antibody), tau (R25863, Zen-Bioscience), tau-pT181 (R23342, Zen-Bioscience), tau-pS396 (381213, Zen-Bioscience), tau-pS262 (310195, Zen-Bioscience), PP2Ac (R25422, Zen-Bioscience), PP2A-pY307 (380708, Zen-Bioscience), GSK-3β-pS9 (5558, Cell Signaling Technology), APP (R22718, Zen-Bioscience), ADAM10 (A10438, Abclonal), BACE1 (A11533, Abclonal), PSEN1 (A19103, Abclonal), Synaptophysin (sc-17750, Santa Cruz Biotechnology), PSD95 (MABN68, Merck Millipore), Map2 (ab254264, Abcam), β-actin (AC026, Abclonal).

    Techniques: De-Phosphorylation Assay, Activity Assay, Western Blot, Inhibition, Molecular Weight

    2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and tau-pS396 in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.

    Journal: Aging (Albany NY)

    Article Title: Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets

    doi: 10.18632/aging.204336

    Figure Lengend Snippet: 2JY-OBZ4 resisted tau hyperphosphorylation in HEK293-hTau cells transfected with GSK-3β plasmid. ( A ) Western blots and ( B – D ) quantitative analysis for GSK-3β, Tau, tau-pT181 and tau-pS396 in HEK293-hTau cells overexpressed with GSK-3β. MW Molecular weight. n = 3 per group. NS means no significance, p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05 and ##p < 0.01.

    Article Snippet: The primary antibodies utilized in this study are as follows: GSK-3β (21002, Signalway Antibody), tau (R25863, Zen-Bioscience), tau-pT181 (R23342, Zen-Bioscience), tau-pS396 (381213, Zen-Bioscience), tau-pS262 (310195, Zen-Bioscience), PP2Ac (R25422, Zen-Bioscience), PP2A-pY307 (380708, Zen-Bioscience), GSK-3β-pS9 (5558, Cell Signaling Technology), APP (R22718, Zen-Bioscience), ADAM10 (A10438, Abclonal), BACE1 (A11533, Abclonal), PSEN1 (A19103, Abclonal), Synaptophysin (sc-17750, Santa Cruz Biotechnology), PSD95 (MABN68, Merck Millipore), Map2 (ab254264, Abcam), β-actin (AC026, Abclonal).

    Techniques: Transfection, Plasmid Preparation, Western Blot, Molecular Weight

    2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Journal: Aging (Albany NY)

    Article Title: Novel small molecular compound 2JY-OBZ4 alleviates AD pathology in cell models via regulating multiple targets

    doi: 10.18632/aging.204336

    Figure Lengend Snippet: 2JY-OBZ4 induced tau dephosphorylation in HEK293/tau cells via upregulating the activity of PP2A. ( A ) Western blots and ( B – G ) quantitative analysis for Tau-pT181, Tau-pS262, Tau-pS396, Tau, PP2Ac, PP2Ac-pY307 (PP2Ac phosphor-Tyr 307 , an indicator of inhibition of PP2A), GSK-3β, GSK-3β-pS9 (GSK-3β phosphor-Ser 9 , an indicator of inhibition of GSK-3β) in HEK293-hTau cells. PP2Ac represents catalytic subunit of PP2A holoenzyme. MW Molecular weight. n = 3 per group. ( H ) PP2A activity was detected in HEK293-hTau cells. n = 4 per group. ( I ) GSK-3β activity was detected in HEK293-hTau cells. n = 3 per group. p value significance is calculated from a one-way ANOVA, data are represented as mean ± SEM. *p < 0.05, ** p < 0.01, **** p < 0.0001, compared to controls and # p < 0.05.

    Article Snippet: The primary antibodies utilized in this study are as follows: GSK-3β (21002, Signalway Antibody), tau (R25863, Zen-Bioscience), tau-pT181 (R23342, Zen-Bioscience), tau-pS396 (381213, Zen-Bioscience), tau-pS262 (310195, Zen-Bioscience), PP2Ac (R25422, Zen-Bioscience), PP2A-pY307 (380708, Zen-Bioscience), GSK-3β-pS9 (5558, Cell Signaling Technology), APP (R22718, Zen-Bioscience), ADAM10 (A10438, Abclonal), BACE1 (A11533, Abclonal), PSEN1 (A19103, Abclonal), Synaptophysin (sc-17750, Santa Cruz Biotechnology), PSD95 (MABN68, Merck Millipore), Map2 (ab254264, Abcam), β-actin (AC026, Abclonal).

    Techniques: De-Phosphorylation Assay, Activity Assay, Western Blot, Inhibition, Molecular Weight