gst keima cb5 Search Results


prk5 gst keima cb5  (Addgene inc)
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Addgene inc prk5 gst keima cb5
A. Western blot showing levels of the indicated proteins in non-PDAC (HEK293T and Hela) and PDAC (KP4, PaTu8902, Panc10.05, PaTu8988T) cells. B. Immunoblot for total levels of FLAG and Tapasin (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in the indicated cell lines. Cells were pre-treated with 400 nM Bafilomycin A1 (BafA1) for 6 h to block degradation. C. Time course measurement of amino acid starvation (EBSS) induced ER-phagy flux in the indicated cell lines stably expressing <t>Keima-cb5</t> relative to DMEM growth media. N = 6 independent experiments. P values were determined using a two-way analysis of variance. D. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, - B, -C) in KP4 cells upon amino acid starvation in EBSS for 9 or 22 h compared to DMEM. N = 3 independent experiments. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. E. Immunoblot for the indicated total protein (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in KP4 cells under full nutrient conditions (DMEM), treatment with 400 nM BafA1 for 6 h, amino acid (AA) starvation for 6 h (EBSS), or combined AA starvation and 400nM BafA1 treatment. Blots are representative of N = 3 independent experiments. F. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, -B, -C) after siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1 in KP4 cells. N = 3 independent experiment. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. G. Immunoblot of the indicated proteins in KP4 cells following siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1. Blots are representative of N = 3 independent experiments. Quantification is relative to siCTRL. H. Immunofluorescence staining of KP4 cells for TEX264 (green), MHC-I (red), and DAPI (blue). Inset shows the magnified view of the region indicated by the white box. Scale bar: 10 µm.
Prk5 Gst Keima Cb5, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/gst+keima+cb5/GST-Keima-cb5+(Plasmid+%23137755)/bio_rxiv__2024__10__27__620516-183-61-68
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prk5 gst keima cb5 - by Bioz Stars, 2026-09
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A. Western blot showing levels of the indicated proteins in non-PDAC (HEK293T and Hela) and PDAC (KP4, PaTu8902, Panc10.05, PaTu8988T) cells. B. Immunoblot for total levels of FLAG and Tapasin (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in the indicated cell lines. Cells were pre-treated with 400 nM Bafilomycin A1 (BafA1) for 6 h to block degradation. C. Time course measurement of amino acid starvation (EBSS) induced ER-phagy flux in the indicated cell lines stably expressing Keima-cb5 relative to DMEM growth media. N = 6 independent experiments. P values were determined using a two-way analysis of variance. D. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, - B, -C) in KP4 cells upon amino acid starvation in EBSS for 9 or 22 h compared to DMEM. N = 3 independent experiments. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. E. Immunoblot for the indicated total protein (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in KP4 cells under full nutrient conditions (DMEM), treatment with 400 nM BafA1 for 6 h, amino acid (AA) starvation for 6 h (EBSS), or combined AA starvation and 400nM BafA1 treatment. Blots are representative of N = 3 independent experiments. F. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, -B, -C) after siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1 in KP4 cells. N = 3 independent experiment. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. G. Immunoblot of the indicated proteins in KP4 cells following siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1. Blots are representative of N = 3 independent experiments. Quantification is relative to siCTRL. H. Immunofluorescence staining of KP4 cells for TEX264 (green), MHC-I (red), and DAPI (blue). Inset shows the magnified view of the region indicated by the white box. Scale bar: 10 µm.

Journal: bioRxiv

Article Title: A multi-subunit autophagic capture complex facilitates degradation of ER stalled MHC-I in pancreatic cancer

doi: 10.1101/2024.10.27.620516

Figure Lengend Snippet: A. Western blot showing levels of the indicated proteins in non-PDAC (HEK293T and Hela) and PDAC (KP4, PaTu8902, Panc10.05, PaTu8988T) cells. B. Immunoblot for total levels of FLAG and Tapasin (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in the indicated cell lines. Cells were pre-treated with 400 nM Bafilomycin A1 (BafA1) for 6 h to block degradation. C. Time course measurement of amino acid starvation (EBSS) induced ER-phagy flux in the indicated cell lines stably expressing Keima-cb5 relative to DMEM growth media. N = 6 independent experiments. P values were determined using a two-way analysis of variance. D. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, - B, -C) in KP4 cells upon amino acid starvation in EBSS for 9 or 22 h compared to DMEM. N = 3 independent experiments. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. E. Immunoblot for the indicated total protein (input) and following immunoprecipitation (IP) of stably expressed HLA-A-FLAG in KP4 cells under full nutrient conditions (DMEM), treatment with 400 nM BafA1 for 6 h, amino acid (AA) starvation for 6 h (EBSS), or combined AA starvation and 400nM BafA1 treatment. Blots are representative of N = 3 independent experiments. F. Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, -B, -C) after siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1 in KP4 cells. N = 3 independent experiment. Data are the mean ± S.D. P values were determined using a one-way analysis of variance. G. Immunoblot of the indicated proteins in KP4 cells following siRNA-mediated knockdown of TEX264, FAM134B, and CCPG1. Blots are representative of N = 3 independent experiments. Quantification is relative to siCTRL. H. Immunofluorescence staining of KP4 cells for TEX264 (green), MHC-I (red), and DAPI (blue). Inset shows the magnified view of the region indicated by the white box. Scale bar: 10 µm.

Article Snippet: pLJM1-HLA-A-FLAG was generated by subcloning the pcDNA of HLA-A (Addgene #85162) together with 2× FLAG into the EcoRI and NheI sites of the pLJM1 lentiviral vector. pMXs-GFP-NBR1, pMXs-GFP-NBR1 ΔUBA, and pMXs-GFP-NBR1 ΔLIR were provided by J. Debnath. pMRX-INU-TEX264-FLAG and pMRX-INU-TEX264 LIR4A- FLAG were gifts from Noboru Mizushima (Addgene #128258 and #128259). pLJM1- Keima-cb5 was generated by cloning the Keima-cb5 fragment from pRK5-GST-Keima- cb5 (a gift from Carol Mercer; Addgene plasmid #137755) into the AgeI and EcoRI sites of the pLJM1 lentiviral vector. pHAGE-CMV-ENV-SAV10-GFP and pHAGE-CMV-ENV- GFP were developed by Peter M. Bruno.

Techniques: Western Blot, Immunoprecipitation, Stable Transfection, Blocking Assay, Expressing, Flow Cytometry, Membrane, Knockdown, Immunofluorescence, Staining

A. Flow cytometry gating strategy for Keima-cb5 experiments shown in . B,C . Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, -B, -C) in PaTu8988T ( B ) or Panc1 ( C ) cells following siRNA-mediated knockdown of TEX264, FAM138B, or CCPG1. N = 3 independent experiments. Data are the mean ± S.D. P values were determined using a one-way analysis of variance.

Journal: bioRxiv

Article Title: A multi-subunit autophagic capture complex facilitates degradation of ER stalled MHC-I in pancreatic cancer

doi: 10.1101/2024.10.27.620516

Figure Lengend Snippet: A. Flow cytometry gating strategy for Keima-cb5 experiments shown in . B,C . Flow cytometry-based quantification of plasma membrane levels of MHC-I (HLA-A, -B, -C) in PaTu8988T ( B ) or Panc1 ( C ) cells following siRNA-mediated knockdown of TEX264, FAM138B, or CCPG1. N = 3 independent experiments. Data are the mean ± S.D. P values were determined using a one-way analysis of variance.

Article Snippet: pLJM1-HLA-A-FLAG was generated by subcloning the pcDNA of HLA-A (Addgene #85162) together with 2× FLAG into the EcoRI and NheI sites of the pLJM1 lentiviral vector. pMXs-GFP-NBR1, pMXs-GFP-NBR1 ΔUBA, and pMXs-GFP-NBR1 ΔLIR were provided by J. Debnath. pMRX-INU-TEX264-FLAG and pMRX-INU-TEX264 LIR4A- FLAG were gifts from Noboru Mizushima (Addgene #128258 and #128259). pLJM1- Keima-cb5 was generated by cloning the Keima-cb5 fragment from pRK5-GST-Keima- cb5 (a gift from Carol Mercer; Addgene plasmid #137755) into the AgeI and EcoRI sites of the pLJM1 lentiviral vector. pHAGE-CMV-ENV-SAV10-GFP and pHAGE-CMV-ENV- GFP were developed by Peter M. Bruno.

Techniques: Flow Cytometry, Membrane, Knockdown