anti phospho p70 ribosomal s6 protein kinase p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70 ribosomal s6 protein kinase p70s6k thr389
    Anti Phospho P70 Ribosomal S6 Protein Kinase P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti phospho thr389 p70 s6 kinase  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho thr389 p70 s6 kinase
    Anti Phospho Thr389 P70 S6 Kinase, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k thr389
    Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k thr389
    TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the <t>mTORC1/p70S6K</t> signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.
    Anti Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Regulation of TSC2 lysosome translocation and mitochondrial turnover by TSC2 acetylation status"

    Article Title: Regulation of TSC2 lysosome translocation and mitochondrial turnover by TSC2 acetylation status

    Journal: Scientific Reports

    doi: 10.1038/s41598-024-63525-7

    TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the mTORC1/p70S6K signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.
    Figure Legend Snippet: TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the mTORC1/p70S6K signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.

    Techniques Used: Membrane, Immunofluorescence, Immunoprecipitation, Western Blot, Negative Control

    anti phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k thr389
    Anti Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k thr389
    a–c qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were stimulated with different doses of pipecolic acid (5, 10, 20 μM) and 100 ng/ml LPS for 6 h. n = 6 per group. d Representative blots and quantified data of the expression of phospho-mTOR (Ser2448), mTOR, <t>phosphop-p70s6k</t> <t>(Thr389),</t> <t>p70s6k,</t> phosphop-S6 (Ser240/244), S6 in the BMDMs. BMDMs were pre-treated with 20 μM pipecolic acid for 1 h before LPS (100 ng/ml, 6 h) stimulation. n = 4 per group. e–g qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were pre-treated with 100 nM rapamycin for 1 h before pipecolic acid (20 μM, 6 h) and LPS (100 ng/ml, 6 h) stimulation. n = 6 per group. Values are presented as mean ± SEM. Data are analyzed using one-way ANOVA followed by Bonferroni’s test. * p < 0.05; ** p < 0.01; *** p < 0.001; exact p values are listed in Source Data file. Pip=pipecolic acid; Rapa=rapamycin. Source data are provided as a Source Data file.
    Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Early-life exercise induces immunometabolic epigenetic modification enhancing anti-inflammatory immunity in middle-aged male mice"

    Article Title: Early-life exercise induces immunometabolic epigenetic modification enhancing anti-inflammatory immunity in middle-aged male mice

    Journal: Nature Communications

    doi: 10.1038/s41467-024-47458-3

    a–c qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were stimulated with different doses of pipecolic acid (5, 10, 20 μM) and 100 ng/ml LPS for 6 h. n = 6 per group. d Representative blots and quantified data of the expression of phospho-mTOR (Ser2448), mTOR, phosphop-p70s6k (Thr389), p70s6k, phosphop-S6 (Ser240/244), S6 in the BMDMs. BMDMs were pre-treated with 20 μM pipecolic acid for 1 h before LPS (100 ng/ml, 6 h) stimulation. n = 4 per group. e–g qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were pre-treated with 100 nM rapamycin for 1 h before pipecolic acid (20 μM, 6 h) and LPS (100 ng/ml, 6 h) stimulation. n = 6 per group. Values are presented as mean ± SEM. Data are analyzed using one-way ANOVA followed by Bonferroni’s test. * p < 0.05; ** p < 0.01; *** p < 0.001; exact p values are listed in Source Data file. Pip=pipecolic acid; Rapa=rapamycin. Source data are provided as a Source Data file.
    Figure Legend Snippet: a–c qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were stimulated with different doses of pipecolic acid (5, 10, 20 μM) and 100 ng/ml LPS for 6 h. n = 6 per group. d Representative blots and quantified data of the expression of phospho-mTOR (Ser2448), mTOR, phosphop-p70s6k (Thr389), p70s6k, phosphop-S6 (Ser240/244), S6 in the BMDMs. BMDMs were pre-treated with 20 μM pipecolic acid for 1 h before LPS (100 ng/ml, 6 h) stimulation. n = 4 per group. e–g qRT-PCR determination of pro-inflammatory cytokines (TNF, IL-1β, and iNOS) mRNA. BMDMs were pre-treated with 100 nM rapamycin for 1 h before pipecolic acid (20 μM, 6 h) and LPS (100 ng/ml, 6 h) stimulation. n = 6 per group. Values are presented as mean ± SEM. Data are analyzed using one-way ANOVA followed by Bonferroni’s test. * p < 0.05; ** p < 0.01; *** p < 0.001; exact p values are listed in Source Data file. Pip=pipecolic acid; Rapa=rapamycin. Source data are provided as a Source Data file.

    Techniques Used: Quantitative RT-PCR, Expressing

    phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k thr389
    Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k thr389
    Anti Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho p70s6k thr389/product/Cell Signaling Technology Inc
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    anti phospho p70s6k thr389 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k thr389 antibody
    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated <t>p70S6k</t> were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.
    Anti Phospho P70s6k Thr389 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho p70s6k thr389 antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    anti phospho p70s6k thr389 antibody - by Bioz Stars, 2024-07
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    1) Product Images from "Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca 2+ -dependent AMPK/mTOR pathway"

    Article Title: Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca 2+ -dependent AMPK/mTOR pathway

    Journal: International Journal of Ophthalmology

    doi: 10.18240/ijo.2024.03.03

    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.
    Figure Legend Snippet: A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.

    Techniques Used: Western Blot, Fluorescence, Flow Cytometry, Expressing

    anti phospho p70s6k thr389 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k thr389 antibody
    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated <t>p70S6k</t> were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.
    Anti Phospho P70s6k Thr389 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho p70s6k thr389 antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p70s6k thr389 antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca 2+ -dependent AMPK/mTOR pathway"

    Article Title: Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca 2+ -dependent AMPK/mTOR pathway

    Journal: International Journal of Ophthalmology

    doi: 10.18240/ijo.2024.03.03

    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.
    Figure Legend Snippet: A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.

    Techniques Used: Western Blot, Fluorescence, Flow Cytometry, Expressing

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    Cell Signaling Technology Inc anti phospho p70 ribosomal s6 protein kinase p70s6k thr389
    Anti Phospho P70 Ribosomal S6 Protein Kinase P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the <t>mTORC1/p70S6K</t> signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.
    Anti Phospho P70s6k Thr389, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated <t>p70S6k</t> were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.
    Anti Phospho P70s6k Thr389 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the mTORC1/p70S6K signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.

    Journal: Scientific Reports

    Article Title: Regulation of TSC2 lysosome translocation and mitochondrial turnover by TSC2 acetylation status

    doi: 10.1038/s41598-024-63525-7

    Figure Lengend Snippet: TSC2 localizes in the lysosomal membrane when it is deacetylated in MEFs. ( A ) Immunofluorescence showing the association between LAMP1 (red channel) and TSC2 (green channel) in MEF TSC2+/+ and Sirt1+/+ cells under control conditions and after resveratrol treatment at 50 µM. The graphs represent the Manders’ colocalization coefficient M1 of TSC2-LAMP1 in either MEF Tsc2+/+ and MEF Sirt1+/+. The values represent means and SD; n = 3 independent experiments. *p ≤ 0.05 comparing MEF Tsc2+/+ control versus resveratrol treatment and MEF Sirt1+/+ control and after the addition of resveratrol. Bars, 20 μm. ( B ) Immunoprecipitation of TSC2 and western blotting against LAMP2A under normal conditions and after the treatment with resveratrol at 50 µM in either MEF Tsc2+/+, Tsc2−/−, Sirt1+/+ and Sirt1−/−. It is shown in the last lane a negative control using a non-specific antibody for the immunoprecipitation. It is also shown in the whole cell extracts (WCE) the mTORC1/p70S6K signalling pathway with the corresponding quantification and statistical analysis. ( C , D ) Representative western blots corresponding to the mTORC1 and AMPK signalling pathways using either nicotinamide (5 mM), resveratrol (50 µM) or control conditions either comparing MEF Tsc2+/+ and −/− cells ( C ) or MEF Sirt1+/+ and −/− cells in ( D ). It is shown in the graphs (in C and in D ) the densitometry corresponding to the P-ACC/ACC ratio in all the cell lines analyzed. The values represent means and SD; n = 6. **p ≤ 0.01, ***p ≤ 0.001 MEF Tsc2+/+ and Sirt1+/+ vs MEF Tsc2−/− and Sirt1−/−.

    Article Snippet: The following antibodies were obtained from Cell Signaling Technology (Beverly, MA): anti-LC3B (#4108), anti-acetyl lysine (#9441), anti-p70S6K (#9202), anti-phospho-p70S6K (Thr389) (#9205), anti-TSC2 (D93F12, #4308), anti-cleaved caspase-3 (#9661), anti-phospho-ACC (Ser 79) (#3661), anti-LAMP1 (#15665), anti-Rheb (#13879), anti-PINK1 (#6946) and anti-phospho-Drp1 (Ser 616) (#3455).

    Techniques: Membrane, Immunofluorescence, Immunoprecipitation, Western Blot, Negative Control

    A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.

    Journal: International Journal of Ophthalmology

    Article Title: Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca 2+ -dependent AMPK/mTOR pathway

    doi: 10.18240/ijo.2024.03.03

    Figure Lengend Snippet: A: The effects of hyperosmotic stress on the levels of phosphorylated mTOR were assessed using Western blotting at 6h. B: Quantitative analysis of the results shown in A. C: The effects of hyperosmotic stress on the levels of phosphorylated AMPK were detected using Western blotting at 6h. D: Quantitative analysis of the data presented in C. E: The effects of hyperosmotic stress on the levels of phosphorylated p70S6k were detected using Western blotting at 6h. F: Quantitative analysis of the data presented in E. G: The effects of BAPTA-AM on the relative conversion of LC3B-II to LC3B-I in LECs treated with hyperosmotic stress were detected using Western blotting at 6h. H: Quantitative analysis of the data presented in G. I-J: The effects of hyperosmotic shock on intracellular Ca2+ levels were detected using a fluorescence microplate reader and flow cytometry. K: The TRPV1 protein expression at 0, 5min, 10min, 15min and 30min after 600 mOsm Treatment. The solid black triangle refers to the first and second band which are qualified the amount of TRPV1. The solid five-pointed star refers to the third band which is considered as degradation. The protein expression of TRPV1 increased mostly after 5min treatment by 600 mOsm. aP<0.05, bP<0.01, dP<0.0001 indicate significant differences from the control; ns indicates no significant difference. All experiments were repeated at least three times. AMPK: AMP-activated protein kinase; mTOR: Mammalian target of rapamycin; BAPTA-AM: Bis-(o-aminophenoxy) ethane-N, N, N′, N′-tetraacetic acid, tetra(acetoxymethyl) ester; LC3: Microtubule-associated protein 1 light chain 3; LEC: Lens epithelial cell; TRPV 1: Transient receptor potential vanilloid 1.

    Article Snippet: The primary antibodies were as follows: anti-transient receptor potential vanilloid 1 (TRPV1) antibody (Alomone labs, # ACC-030, rabbit), anti-LC3B antibody (CST, #2775, 1:1000, rabbit), anti-LC3B antibody (E5Q2K; CST, #83506, 1:1000, mouse), anti-Beclin 1 antibody (D40C5; CST, #3495, 1:1000, rabbit), anti-sequestosome-1 (SQSTM1) antibody (Proteintech, #18420-1-AP, 1:1000, rabbit), anti-autophagy-related gene (ATG) 7 (D12B11; CST, #8558, 1:1000, rabbit), anti-p70S6K antibody (CST, #9202, 1:1000, rabbit), anti-phospho-p70S6K (Thr389) antibody (CST, #9234, 1:1000, rabbit), anti-mammalian target of rapamycin (mTOR) antibody (CST, #2972, 1:1000, rabbit), anti-phospho-mTOR antibody (Ser2448; CST, #2971, 1:1000, rabbit), anti-AMP-activated protein kinase (AMPK) α antibody (CST, #2532, 1:1000, rabbit), anti-phospho-AMPKα (Thr172) antibody (CST, #2535, 1:1000, rabbit), and anti-β-actin antibody (CST, #4970, 1:5000, rabbit or CST, #3700, 1:5000, mouse).

    Techniques: Western Blot, Fluorescence, Flow Cytometry, Expressing