phospho p70s6k  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k
    AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase <t>(p70S6K),</t> phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.
    Phospho P70s6k, 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 "Direct and acute effects of advanced glycation end products on proteostasis in isolated mouse skeletal muscle"

    Article Title: Direct and acute effects of advanced glycation end products on proteostasis in isolated mouse skeletal muscle

    Journal: Physiological Reports

    doi: 10.14814/phy2.16121

    AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase (p70S6K), phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.
    Figure Legend Snippet: AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase (p70S6K), phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.

    Techniques Used: Western Blot, Binding Assay

    phospho p70s6k  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k
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    phospho p70s6k thr389  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k thr389
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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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    anti phospho p70s6k  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k
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    anti phospho p70s6k  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p70s6k
    The mTOR signaling pathway participates in the damage to the BSCB damage following EC dysfunction. A , B , C Protein levels of p-mTOR, <t>p-P70S6K,</t> p-AMPK, p-4EBP1 detected by Western blot (left) and their quantitative analyses (right). * p <0.05, ** p <0.01, *** p <0.001
    Anti Phospho P70s6k, 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 "Glucose competition between endothelial cells in the blood-spinal cord barrier and infiltrating regulatory T cells is linked to sleep restriction-induced hyperalgesia"

    Article Title: Glucose competition between endothelial cells in the blood-spinal cord barrier and infiltrating regulatory T cells is linked to sleep restriction-induced hyperalgesia

    Journal: BMC Medicine

    doi: 10.1186/s12916-024-03413-z

    The mTOR signaling pathway participates in the damage to the BSCB damage following EC dysfunction. A , B , C Protein levels of p-mTOR, p-P70S6K, p-AMPK, p-4EBP1 detected by Western blot (left) and their quantitative analyses (right). * p <0.05, ** p <0.01, *** p <0.001
    Figure Legend Snippet: The mTOR signaling pathway participates in the damage to the BSCB damage following EC dysfunction. A , B , C Protein levels of p-mTOR, p-P70S6K, p-AMPK, p-4EBP1 detected by Western blot (left) and their quantitative analyses (right). * p <0.05, ** p <0.01, *** p <0.001

    Techniques Used: Western Blot

    phospho p70s6k  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p70s6k
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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.
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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
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    Cell Signaling Technology Inc phospho p70s6k
    AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase <t>(p70S6K),</t> phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.
    Phospho P70s6k, 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/phospho p70s6k/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc phospho p70s6k thr389
    AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase <t>(p70S6K),</t> phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.
    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/phospho p70s6k thr389/product/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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    Cell Signaling Technology Inc anti phospho p70s6k
    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, 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 - by Bioz Stars, 2024-06
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    AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase (p70S6K), phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.

    Journal: Physiological Reports

    Article Title: Direct and acute effects of advanced glycation end products on proteostasis in isolated mouse skeletal muscle

    doi: 10.14814/phy2.16121

    Figure Lengend Snippet: AGEs stimulation reduces the protein synthesis pathway. (a) Representative immunoblots from western blotting. (b) Total mechanistic target of rapamycin (mTOR), phosphorylation of mTOR Ser 2448 , the ratio of phosphorylation to total mTOR (6 h of AGEs vs. 6 h of BSA, g = 1.34; vs. 2 h of AGEs, g = 1.75). (c) Total 70 kDa ribosomal protein S6 kinase (p70S6K), phosphorylation of p70S6K Thr 389 , the ratio of phosphorylation to total p70S6K. (d) Total 4E binding protein 1 (4E‐BP1), phosphorylation of 4E‐BP1 Thr 37/46 , the ratio of phosphorylation to total 4E‐BP1. Data are shown as box plots. Individual data points are indicated on the graph. Statistical significance is analyzed using two‐way ANOVA with simple effects tests. n = 9/group.

    Article Snippet: The following are the data of the primary antibody: puromycin (Millipore, MABE343), phospho‐p70S6K (Cell Signaling Technology [CST], Danvers, MA, USA, 9234), p70S6K (CST, 9202), phospho‐mTOR (CST, 2971), mTOR (CST, 2972), phospho‐4E‐BP1 (CST, 9459), 4E‐BP1 (CST, 9452), p62 (CST, 5114), LC3B (CST, 2775), MuRF (Santa Cruz, Santa Cruz, CA, USA, sc‐398,609), Atrogin‐1 (Santa Cruz, sc‐27,645), CHOP (CST, 2895), ATF6 (Gene Tex, Arling‐ton, TX, USA, GTX104820), IRE1α (CST, 3924), phospho‐IRE1 (Abcam, ab48187), PERK (CST, 5683), phospho‐PERK (ABclonal, Woburn, MA, USA AP0886), XBP‐1 s (CST, 40435), OXPHOS (Abcam, Cambridge, UK, ab110413), PGC‐1α (Abcam, ab54481), p38MAPK (CST, 9212), phospho‐p38 MAPK (CST, 9211), SAP/JNK (CST, 9252), phospho‐SAP/JNK (CST, 9251), p44/42 MAPK (Erk1/2) (CST, 9102), and phospho‐p44/42 MAPK (Erk1/2) (CST, 9101).

    Techniques: Western Blot, Binding Assay

    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