This readme.txt file was generated on [2019-01-16] by [Dan Mulvihill] 
GENERAL INFORMATION 
1.
 Title of Dataset 
TORC2 dependent phosphorylation modulates calcium regulation of fission yeast myosin. 


2.
 Author Information Principal Investigator Contact Information


Name: Dr. Daniel P. Mulvihill
 Institution: University of Kent
Address: School of Biosciences, Canterbury, Kent, CT27NJ, UK
Email: d.p.mulvihill@kent.ac.uk

 Associate or Co-investigators Contact Information
Name: Dr. Karen Baker
 Institution: University of Kent
Address: School of Biosciences, Canterbury, Kent, CT27NJ, UK
Email: k.b.baker@kent.ac.uk

 Name: Ms. Irene Gyamfi
Institution: University of Kent
Address: School of Biosciences, Canterbury, Kent, CT27NJ, UK
Email: iag7@kent.ac.uk

 Name: Prof. Michael Geeves 
Institution: University of Kent
Address: School of Biosciences, Canterbury, Kent, CT27NJ, UK
Email: m.a.geeves@kent.ac.uk

 Name: Dr. Gregory Mashanov
Institution: The Francis Crick Institute 
Address:1 Midland Road, London NW1 1AT, UK
Email: gregory.mashanov@crick.ac.uk 

Name: Dr. Justin Molloy
Institution: The Francis Crick Institute 
Address:1 Midland Road, London NW1 1AT, UK
Email: justin.molloy@crick.ac.uk 

3.
 Date of data collection: 2010 to 2018 

4.
 Geographic location of data collection (where was data collected?):


-School of Biosciences, University of Kent, Canterbury.
-National Institute for Medical Research, The Ridgeway, London NW7 1AA.
-The Francis Crick Institute, Address:1 Midland Road, London NW1 1AT, UK 5. Information about funding sources that supported the collection of the data:This work was supported by the University of Kent and funding from the Biotechnology and BiologicalSciences Research Council (BB/J012793/1 & BB/M015130/1), a Royal Society Industry Fellowship to DPM; aCASE industrial bursary from Cairn Research Ltd to KB and by the Francis Crick Institute which receives corefunding from Cancer Research UK (FC001119), the UK Medical Research Council (FC001119) and theWellcome Trust (FC001119) GIM and JEM. 
DATA & FILE OVERVIEW 
1. File List 
Short description: Compressed ZIP files of

-Widefield (WF) live cell imaging data

-Total Internal Reflection Fluorescence (TIRF) live cell imaging data

-Western Blot Scans 

-Biochemical data 

-Data Analysis Spreadsheets 

2. Relationship between files:
ZIP files contain raw data associated with this manuscript.

-Biochemistry data files: Contain Origin files containing raw data and analysis.

-Western Blot Original Scans: Contains JPEG files of original scans of western blot membranes shown in
paper. Designation at beginning of file name denotes figure component within paper.

-Widefield data files ('WF' at beginning of ZIP files): Contain .TIFF files showing Myo1, Cam1, Cam2 or Sla2
localisation and dynamics in wild type, myo1.S742A,myo1D or cam2D backgrounds.

-Total Internal Reflection Fluorescence ('TIRF' at beginning of ZIP files): Contain .gmv files of Myo1, Cam1
or Cam2 localisation and dynamics in wild type or myo1.S742A backgrounds.

-Data Analysis Files: Excel spreadsheets showing image intensities and dynamics of Myo1, Cam1, Cam2,
Sla2 foci within WF and TIRF data sets, measured using Metamorph, Autoquant, or GMimPro software. 

3.
 Additional related data collected that was not included in the current data package:
None 


4.
 Are there multiple versions of the dataset?
No 



METHODOLOGICAL INFORMATION 
1. Description of methods used for collection/generation of data:Widfield imaging: Samples were visualized using an Olympus IX71 microscope with PlanApo 100x OTIRFM­SP 1.45 NA lens mounted on a PIFOC z-axis focus drive (Physik Instrumente, Karlsruhe, Germany), andilluminated using LED light sources (Cairn Research Ltd, Faversham, UK) with appropriate filters (Chroma,Bellows Falls, VT). Samples were visualized using a QuantEM
(Photometrics) EMCCD camera, and the system was controlled with
Metamorph software (Molecular Devices). 3D dataset files for fluorescent images are composed of 13 x z-plane images were each 0.2 µm apart. During live-cell imaging, cells were mounted onto coverslips withlectin (Sigma L2380; 1 mg/ml) in a Bioptechs FCS2 (Bioptechs, Butler, PA), fitted onto an ASI motorizedstage (ASI, Eugene, OR) on the above system, with the sample holder, objective lens and environmental chamber held at the required temperature. All live-cell imaging was undertaken using EMMG media. 
2.
 Methods for processing the data:
File contains unprocessed raw data files. 


3.
 Instrument- or software-specific information needed to interpret the data:
Widefield data acquired using MDA Metamorph software. Can be visualised using variety of standard image
analysis packages. Freely available open source software packages available, such as Fiji
(https://imagej.net/ImageJ) or Micromanager (https://micro-manager.org/). 


TIRF data acquired using the open source -GMimPro software- package. 

4.
 Standards and calibration information, if appropriate: 


5.
 Environmental/experimental conditions:
Cells were mounted directly from the culture (without centrifugation) onto lectin-coated coverslips, bathed in



EMMG media and imaged at 20 C. 
6.
 Describe any quality-assurance procedures performed on the data: -

7.
 People involved with sample collection, processing, analysis and/or submission: 


Matthew Johnson: Strain generation, cell culture, sample preparation, and image acquisition.Daniel Mulvihill: Manuscript preparation and data submission. 
DATA-SPECIFIC INFORMATION FOR: [Myo1-S742] 
Widefield data: 
1.
 Number of variables: 4 

2.
 Number of cases/rows: N/A 

3.
 Variable List

 (A)
 Name: 	[GFP labelled protein]
Description: [Description of the variable]
Value labels: mNG-myo1, cam1-GFP, cam1-mCherry, cam2-gfp or sla2-mCherry 


(B)
 Name: [Genetic background]
Description: [denotes gene deletion strain examined]
Value labels: WT (no label), myo1-S742A, cam2D, myo1D


 (C)
 Name: [Emission Wavelength Captured]
Description: [Description of Fluorescence protein or light path]
Value labels: GFP, tdTomato or Phase


 (D)
 Name: Timepoint
Description: Timepoint within time-lapse experiment. 





 Value labels: 001,002, etc 
Example filename with variables in brackets: mNeogreen-myo1(A) cam2D(B) 026_w15. tdTomato(C) LED3001(D).TIF 
TIRF data: 
1.
 Number of variables: 1 

2.
 Number of cases/rows: N/A 

3.
 Variable 


Name: Sample number. Strains examined are organised within discrete folders. Each file correlates with dateof experiment, and sample number on the day. 
Example filename with variables in brackets: 050418r01.gmv ( - 1st experiment undertaken on 5th April2018) - Titles of columns within TIRF data analysis spread sheet denotes this source file number of analysedparticle. 
4.
 Missing data codes:Code/symbol Definition Code/symbol Definition 

5.
 Specialised formats of other abbreviations used
D (delta) - strain in which associated gene has been deleted (e.g. cam2D)