Post-processing:
Once SAINTexpress has been run, APOSTL is able to read the resulting "list.txt" file. Fromhere APOSTL does a number of things:
APOSTL calculates NSAF values for each prey based on the average spectra observed for each bait
OPTIONAL: APOSTL calculates the probability of a specific interaction based on prey revalence in the CRAPome
Bubble graphs are generated for each bait where:
- x axis is the natural log of the NSAF values
- y axis is the observed log2 fold change (as compared to control)
- bubble radius is proportional to the average observed spectra
OPTIONAL: bubble color corresponds to the CRAPome probability of a specific interaction in which an 80% cutoff is applied where prey with less than 80% are colored tan
APOSTL queries ConsensusPathDB for protein-protein interactions within your data and then formats the resulting network for simple cytoscape import using the "import network from file" option.
INPUTS:
MaxQuant or Scaffold:
Select Scaffold if using Scaffold output and MaxQunat if using pepetides.txt from MaxQuant software.
List File:
SAINTexpress generated "list.txt" file
Prey File:
SAINT pre-processing generated "prey.txt" file used to run SAINTexpress
Crapome File:
Raw output from Crapome Workflow 1 query (http://www.crapome.org)
Color:
Specify the color of the bubbles within the graph.
If "crapome" is chosen and 'crapome' file is specified, bubbles will be color based on crapome specificity
Use labels:
Adds gene name labels to bubbles within the "zoomed in" graphs
SAINT Score Cutoff:
Choose Saintscore cutoff (between 0-1) to be used for filtering the "zoomed in" graphs (default = 0.8)
Also used for filtering during the generation of the cytoscape network
Species:
Human, mouse, or yeast
Interaction Confidence:
Interaction confidence value (0-1) used to filter the interactions with the ConsensusPathDB database
Suggestions:
- low: 0
- medium: 0.5
- high: 0.7
- very high: 0.9
List of Uniprot IDs to Include:
Text file with one column of uniprot IDs. Only the Uniprot IDs in the file will be used on the resulting bubblebeam.
List of Uniprot IDs to Exclude:
Text file with one column of uniprot IDs. The Uniprot IDs in this list will not be used in generating the bubblebeam.
OUTPUTS:
Bubble Graphs:
Unfiltered Data:
Filtered by SAINT Score
Output.txt
SAINTexpress "list.txt" output with additional columns that were used during the analysis.