Dataset for "Membrane extraction with styrene-maleic acid copolymer results in insulin receptor autophosphorylation in the absence of ligand"

1041 51 15093 1 47950 document 15093 Doctoral_Data_Management_Plan_Kerrie_Morrison.docx application/vnd.openxmlformats-officedocument.wordprocessingml.document 5f5ae08d1c59a8d674358fe7a3a45c70 MD5 446948 2021-07-07 14:08:31 https://researchdata.bath.ac.uk/1041/2/Doctoral_Data_Management_Plan_Kerrie_Morrison.docx 1041 2 2 application/vnd.openxmlformats-officedocument.wordprocessingml.document other en staffonly

Doctoral_Data_Management_Plan_Kerrie_Morrison.docx

dmp 15094 1 47954 document 15094 indexcodes.txt text/plain e9f443068b1d1a61535111fd3b1d9387 MD5 2371 2021-07-07 14:17:55 https://researchdata.bath.ac.uk/1041/3/indexcodes.txt 1041 3 3 text/plain other Generate index codes conversion from other to indexcodes en staffonly

indexcodes.txt

http://eprints.org/relation/isVersionOf https://researchdata.bath.ac.uk/id/document/15093 http://eprints.org/relation/isVolatileVersionOf https://researchdata.bath.ac.uk/id/document/15093 http://eprints.org/relation/isIndexCodesVersionOf https://researchdata.bath.ac.uk/id/document/15093 15723 5 51605 document 15723 For_archive.zip application/zip 6e1aab7b822d0784c03c1cf059ddf439 MD5 192865009 2022-02-26 11:06:10 https://researchdata.bath.ac.uk/1041/4/For_archive.zip 1041 4 4 application/zip other .zip file containing the following items for the paper; 1. all raw .tiff files for western blots used for quantification including MS excel spreadsheet depicting which bands from the experiments were used. 2. all raw .tiff files used for representative blots in each figure, with MS Powerpoint document to show annoted versions. 3. READ_ME.txt file to further describe the data present. 4. Denistometry values, calculations and statistics; collated_InsR_values_for_archive.xlsx en public cc_by

For_archive.zip

data 15724 3 51624 document 15724 READ_ME.txt text/plain ef473155a6dd978a033dadd6f70347c5 MD5 1009 2022-02-28 11:24:17 https://researchdata.bath.ac.uk/1041/5/READ_ME.txt 1041 5 5 text/plain other en public cc_by

READ_ME.txt

documentation 15725 1 51628 document 15725 indexcodes.txt text/plain bedd96ec8e0edb541198079b64a8904b MD5 409 2022-02-28 11:34:57 https://researchdata.bath.ac.uk/1041/6/indexcodes.txt 1041 6 6 text/plain other Generate index codes conversion from other to indexcodes en public

indexcodes.txt

http://eprints.org/relation/isVersionOf https://researchdata.bath.ac.uk/id/document/15724 http://eprints.org/relation/isVolatileVersionOf https://researchdata.bath.ac.uk/id/document/15724 http://eprints.org/relation/isIndexCodesVersionOf https://researchdata.bath.ac.uk/id/document/15724 archive 9042 disk0/00/00/10/41 2022-03-04 13:23:00 2024-07-18 12:32:01 2022-03-04 13:23:00 data_collection show Morrison Kerrie K.A.Morrison@bath.ac.uk 0000-0001-7130-1183 University of Bath FALSE Whitley Paul P.R.Whitley@bath.ac.uk 0000-0001-9796-8488 University of Bath TRUE Koumanov Francoise F.Koumanov@bath.ac.uk 0000-0002-9029-3390 University of Bath FALSE Dataset for "Membrane extraction with styrene-maleic acid copolymer results in insulin receptor autophosphorylation in the absence of ligand" BY0040 BY0100 dept_health dept_chem dept_bio A READ_ME.txt file is included to provide further information of the data present in the .zip file. Folders labelled "Figure_X", where X corresponds to the figure number, contain raw .tiff files used for representative blots in the paper. An additional MS PowerPoint file is included to annotate these images. "SX" describes the supplementary figures. All_blots_for_quantification folder contains all raw .tiff files obtained and included in quantification of densitometry values. Experiments_used_for_each_graph.xlsx highlights the experiments and bands used for each quantification, representated by graphs in the paper. The Excel spreadsheet "collated_InsR_values_for_archive.xlsx" has three tabs; 1. Densitometry_values - holds the raw densitometry values calculated with Image Studio Lite (LI-COR Biosciences®); normalisation to Akt in separate column (L); representative western blots for each sample measured and "fold difference" between basal and + Insulin samples. 2. Calculations - taking the fold difference tables for each experiment, with additional tables calculating fold difference relative to POST SMA Basal (Green) and PRE RIPA Basal (Orange). Averages were taken for each sample across multiple experiments and standard deviation (SD) and standard error mean (SEM) were caculated. 3. Stats - Statistic for each protein of study; InsR, IRS and Akt, across multiple experiments using paired t-tests. The data was collected to establish whether the insulin receptor maintained its native functionality once extracted into polymer nanodiscs using styrene maleic acid (SMA). The data gathered here show the raw .tiff files of the western blots. Differences in protein activity were quantified by comparing changes between total and phospho- antibody detection, using densitometry. Downstream subunits associated with the insulin receptor cascade were also quantified in this way. .zip file containing the following items; 1. all raw .tiff files gathered from western blots used for quantification. A MS excel spreadsheet is also included to indicate which bands from the experiments were used for each quantification. 2. all raw .tiff files used for representative blots in each figure, with MS Powerpoint document to show annoted versions for these images. 3. READ_ME.txt file to further describe the data present. 4. Excel spreadsheet with raw data values from densitometry analysis from western blots, representative western blots for each experiment, calculations from the densitometry values and statistics associated with these values. 2022-03-03 University of Bath public RightsHolder University of Bath Engineering and Physical Sciences Research Council https://doi.org/10.13039/501100000266 EP/L016354/1 EPSRC Centre for Doctoral Training in Sustainable Chemical Technologies Science and Technology Facilities Council https://doi.org/10.13039/501100000271 SA-54 ISIS Studentship Agreement Medical Research Council https://doi.org/10.13039/501100000265 MR/P002927/1 Role of Rab3 in Peripheral Tissue Insulin Resistance Diabetes UK https://doi.org/10.13039/501100000361 19/0005983 Endosomal sorting complexes in insulin-stimulated GLUT4 recycling cent_sus_tech Samples were generated using methods discussed in paper manuscript "Membrane Extraction with Styrene-Maleic Acid Copolymer Results in Insulin Receptor Autophosphorylation in the Absence of Ligand." Samples were separated on 10% SDS polyacrylamide gels and transferred onto nitrocellulose using a semi-dry transfer apparatus, before immunodetection with primary antibodies; either total antibodies or phopho-specific antibodies. Blots were incubated with secondary antibody,HRP, followed by washes. ECL™ Select Western blotting detection reagent was used for detection. Images were acquired with EPI Chemi II darkroom (UVP). Quantification of bands on immunoblots was measured by densitometry using Image Studio Lite (LI-COR Biosciences®). Specific band intensities were normalized to total Akt levels for that sample. The ratios between phospho-specific and total protein band intensities were calculated in the same sample. Data has been deposited and can be opened with Microsoft Excel. en 1 10.15125/BATH-01041 https://doi.org/10.1038/s41598-022-07606-5 pub open Aimed at potential readers of journal manuscript "Membrane Extraction with Styrene-Maleic Acid Copolymer Results in Insulin Receptor Autophosphorylation in the Absence of Ligand." Dataset