rabbit anti hbxip lamtor3 Search Results


1 ap  (Proteintech)
91
Proteintech 1 ap
1 Ap, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+hbxip+lamtor3/HBXIP+Antibody/pmc11876615-19-8-4
Average 91 stars, based on 1 article reviews
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lamtor5  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc lamtor5
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Lamtor5, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 1 article reviews
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anti lamtor4 c7orf59  (Atlas Antibodies)
93
Atlas Antibodies anti lamtor4 c7orf59
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Anti Lamtor4 C7orf59, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti lamtor1 p18  (Atlas Antibodies)
93
Atlas Antibodies anti lamtor1 p18
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Anti Lamtor1 P18, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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lamtor4  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc lamtor4
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Lamtor4, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+hbxip+lamtor3/Phospho-4E-BP1+(Thr37%2F46)+Rabbit+mAb/pm35608036-155-62-63
Average 96 stars, based on 1 article reviews
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rabbit anti loh12cr1  (Atlas Antibodies)
86
Atlas Antibodies rabbit anti loh12cr1
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Loh12cr1, supplied by Atlas Antibodies, 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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rabbit anti snapin  (Proteintech)
93
Proteintech rabbit anti snapin
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Snapin, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti raga  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc rabbit anti raga
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Raga, 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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rabbit anti eea1  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology rabbit anti eea1
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Eea1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti ha  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc rabbit anti ha
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Ha, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti lamtor2  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc rabbit anti lamtor2
Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and <t>Lamtor5</t> are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.
Rabbit Anti Lamtor2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and Lamtor5 are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.

Journal: The Journal of Biological Chemistry

Article Title: The Ragulator complex serves as a substrate-specific mTORC1 scaffold in regulating the nuclear translocation of transcription factor EB

doi: 10.1016/j.jbc.2022.101744

Figure Lengend Snippet: Generation of Ragulator-deficient clone with total mTORC1 activity. A , schematic representation of the findings of this study, depicting the phosphorylation of the TFEB substrate by mTORC1 bound to the Ragulator-Rag scaffold. B , the results of Western blotting studies on ΔRagulator and parental RAW264.7 cells. Lamtor1, Lamtor2, Lamtor3, Lamtor4, and Lamtor5 are components of the Ragulator complex; β-actin served as the internal control; mTOR and Raptor are components of mTORC1; RagC is necessary for the docking of TFEB to Ragulator. C , representative result of Western blotting studies for determining the activity of mTORC1. The control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. The phosphorylation of S6K in these cells was abolished following treatment with the mTOR kinase inhibitor, Torin1, at the indicated concentrations for 1 h. D , the densities of the p-S6K and S6K bands in panel C were quantified. The ratio of densities of the p-S6K band to S6K band is shown. Four independent experiments were performed. Bars indicate the median. Statistical significance was calculated using Mann–Whitney U test. E , the results of Upstream Regulator Analysis, as determined using the web-based software, Ingenuity Pathway Analysis. The pathways with z scores less than −2 are significantly inactivated in the ΔRagulator cells, compared to the RAW264.7 cells. The z score for the TFEB pathway was >2, which indicates that TFEB is significantly activated in the ΔRagulator cells than in the control RAW264.7 cells. F , the sizes of the ΔRagulator cells were larger than those of the parental RAW264.7 cells. The cell sizes of 1 × 10 4 cells were estimated using flow cytometry. Statistical significance was calculated using Welch’s t test. G , Western blotting for determining the phosphorylation of TSC2. Control RAW264.7 cells and ΔRagulator cells were cultured in nutrient-sufficient and serum-supplemented DMEM. Three biological triplicates were examined. H , the densities of the p-TSC2 and the TSC2 bands in panel G were quantified. The ratio of densities of the p-TSC2 band to TSC2 band is shown. Statistical significance was calculated using the Mann–Whitney U test. mTORC1, mammalian target of rapamycin complex 1; TFEB transcription factor EB.

Article Snippet: For Western blotting, the antibodies against S6K (#2708), p-S6K (T389, #9234), Akt (#4691), p-Akt (Ser473, #4060), 4E-BP1 (#9644), p-4E-BP1 (#2855), Lamtor1 (#8975), Lamtor2 (#8145), Lamtor3 (#8168), Lamtor4 (#12284), Lamtor5 (#14633), mTOR (#2983), p-mTOR (Ser2448, #5536), Raptor (#2280), RagC (#9480), Rheb (#13879), TSC2 (#4308), and p-TSC2 (Thr1462, #3617) were purchased from Cell Signaling Technology; while the antibodies against Lamin B (sc-6216), LAMP-1(clone 1D4B) and β-actin (clone AC-15) were purchased from Santa Cruz Biotechnology.

Techniques: Activity Assay, Phospho-proteomics, Western Blot, Control, Cell Culture, MANN-WHITNEY, Software, Flow Cytometry