DOI: 10.5281/zenodo.10843994
title: Can calmodulin bind to lipids of the cytosolic leaflet of plasma membranes?

description: Can calmodulin bind to lipids of the cytosolic leaflet of plasma membranes?:

This data set contains all the experimental raw data, analysis and source files for the final figures reported in the manuscript: "Can calmodulin bind to lipids of the cytosolic leaflet of plasma membranes?". It is divided into five (1-5) zipped folders, named as the technique used to obtain the data. Each of them, where applicable, consists of three different subfolders (raw data, analysed data, final graph). Read below for more details.  

1) ConfocalMicroscopy

   1a) Raw_Data: the raw images are reported as .dat and .tif formats, divided into folders (according to date first yymmdd, and within the same day according to composition). Each folder contains a .txt file reporting the experimental details   

   1b) GUVs_Statistics       - GUVs_Statistics.txt explains how we generated the bar plot shown in Fig. 1E

   1c) Final_Graph       - Figure_1B_1D.png is the figure representing figure 1B and 1D       - Figure1E_%ofGUVswithCaMAdsorbptions.csv is the source file x-y of the bar plot shown in figure 1E (% of GUVs which showed adsorption of CaM over the total amount of measured GUVs)        - Where_To_Find_Representative_Images.txt states the folders where the raw images chosen for figure 1 can be found 

2) FCS      2a) Raw_Data:        - 1_points: .ptu files        - 2_points: .ht3 files        - Raw_Data_Description.docx which compositions and conditions correspond to which point in the two data sets      2b) Final_Graphs:       - Figure_2A.xlsx contains the x-y source file for figure 2A

   2c) Analysis:        - FCS_Fits.xlsx outcome of the global fitting procedure described in the .docx below (each group of points represents a certain composition and calcium concentration, read the Raw_Data_Description.docx in the FCS > Raw_Data)       - Notes_for_FCS_Analysis.docx contains a brief description of the analysis of the autocorrelation curves

3) GPLaurdan      3a) Raw Data: all the spectra are stored in folders named by date (yymmdd_lipidcomposition_Laurdan) and are in both .FS and .txt formats 

   3b) GP calculations: contains all the .xlsx files calculating the GP values from the raw emission and excitation spectra

   3c)  Final_Graphs       - Data_Processing_For_Fig_2D.csv contains the data processing from the GP values calculated from the spectra to the DeltaGP (GP with- GP without CaM) reported in fig. 2D       - Figure_2C_2D.xlsx contains the x-y source file for the figure 2C and 2D

4) LiveCellsImaging 

   3a) Intensity_Protrusions_vs_Cell_Body:        - contains all the .xlsx files calculating the intensity of the various images. File renamed by date (yymmdd)        - All data in all excel sheets gathered in another Excel file to create a final graph      

   3b)  Final_Graphs       - Figure_S2B.xlsx contains the x-y source file for the figure S2B

5) LiveCellImaging_Raw_Data: it contains some of the images, which are given in .tif. They are divided by date (yymmdd) and each contains subfolders renamed by sample name, concentration of ionomycin. Within the subfolders, the images are divided into folders distinguishing the data acquired before and after the ionomycin treatment and the incubation time.

 

6)  211124_BioCev_Imaging_1 folder has the .jpg files of the time laps, these are shown in fig 1A and S2.

7) 211124_BioCev_Imaging_2 and 8) 211124_BioCev_Imaging_3 contain the images of HeLa cells expressing EGFP-CaM after treatment with ionomycin 200 nM (A1) and 1 uM (A2), respectively. 

9) SPR

   9a) Raw Data:        - SPR_Raw_Data.xlsx x/y exported sensorgrams        - the .jpg files of the software are also reported and named by lipid composition

   9b) Final_Graph:        - Fig.2B.xlsx contains the x-y source file for the figure 2B

   9c)  Analysis       - SPR_Analysis.xlsx: excel file containing step-by-step (sheet by sheet) how we processed the raw data to obtain the final figure (details explained in the .docx below)       - Analysis of SPR data_notes.docx: read me for detailed explanation