and just click the tabs for some magic!

1. Select or search for your species.

Analyze public RNA-seq datasets for 9 species
?

Loading R packages ... ... ...

Less than 5% users sent us emails of support. We are struggling to get funding to maintain and improve iDEP. Reviewers said iDEP is not useful and other tools exist. iDEP is designed to help small labs make sense of transcriptomes. Please send us a brief email today to support our grant proposal, indicating your broad area of research and how iDEP helps your work. All letters count, even from graduate students in another country. Thanks! Email

New! Massively upgraded annotation database! V0.90 includes 315 organisms in Ensembl release 96, plus all species from STRINGdb (v10):115 archaeal, 1678 bacterial, and 238 eukaryotic species

Now published on BMC Bioinformatics!
Due to lack of funding, iDEP has not been thoroughly tested. Please let us know if you find any issue/bug.
We are happy to help prepare your data for iDEP. Dr. Ge is also open to bioinformatics consulting during the summer.

Only keep genes above this level in at least n samples:
Keep genes with minimal counts per million (CPM) in at least n libraries:
Pseudo count c:




Processed data High-resolution figure

?
Aspect ratios of figures can be adjusted by changing the width of browser window.



Customize hierarchical clustering (Default values work well):
Color
Distance
Linkage
Cut-off Z score
Heatmap data High-resolution figure
?

K-means data High-resolution figure
Pathway database
Enrichment details
?

Enriched pathways for each cluster


Coordinates High-resolution figure
?




Identifying Differential Expressed Genes (DEGs). See next tab for details.
Using the limma package



Gene lists

FDR & fold-changes for all genes
Figure

?


Numbers of differentially expressed genes for all comparisons. "B-A" means B vs. A. Interaction terms start with "I:"

Examine the results of DEGs for each comparison

Gene list & data High-resolution figure
Enrichment analysis for DEGs:
Enrichment details
Also try ShinyGO
?

Enriched pathways in DEGs for the selected comparison:

Top Genes for selected comparison:

Pathway list w/ genes
* Warning! The many combinations can lead to false positives in pathway analyses.
?
Red and blue indicates activated and suppressed pathways, respectively.
Red and blue indicates activated and suppressed pathways, respectively.
Red and green represent up- and down-regulated genes, respectively.
This interactive map shows DEGs on the genome. Red and blue dots represent up- or down-regulated genes, respectively. Mouse over to see gene symbols. Click and drag to zoom in.

To identify genomic regions significatly enriched with up- or down-regulated genes, we can use PREDA. Very slow (5 mins), but may be useful in studying cancer or other diseases that might involve chromosomal gain or loss.
Biclustering can discover genes correlated on subset of samples. Only useful when sample size is large(>10). Uses methods implemented in the biclust R package.
Enrichment database
Download all biclusters

?

Enriched gene sets in selected bicluster



Genes in this cluster

Identify co-expression networks and sub-modules using WGCNA. Only useful when sample size is large(>15).
Enrichment database:
Download all modules Download network for selected module
The network file can be imported to VisANT or Cytoscape.
?


Enriched pathways among all genes in selected module

Email us for questions, suggestions, or data contributions. Stay connected via user group or Twitter. Visit our GitHub page to see source code, install a local version, or report bugs and request features. iDEP is being developed by a very small team: Dr. Xijin Ge and a graduate student (Homepage).

R as in Reproducibility

Documentation site. Source code on GitHub, where users can also report bugs or requet features.
To improve reproducibility, iDEP generates custom R code based on your data and choices of parameters. Users with some R coding experience should be able to re-run most analyses by downloading all of the files below. If Ensembl IDs is not used in users' original file, we should use the converted data file. Click through all the tabs and then download all these file to a folder. Run the Customized R code or the Markdown file. R Markdown example.
Customized R code Customized R code(Markdown) iDEP core functions Gene Info file Pathway file (large)

Previous versions of iDEP

iDEP 0.85 with Ensembl 95, archived on May 19, 2019
iDEP 0.82 with Ensembl BioMart version 92, archived on March 29, 2019
iDEP 0.73 with Ensembl BioMart version 91, archived on July 11, 2018

Citation

Please cite: Ge SX, Son EW, Yao R: iDEP: an integrated web application for differential expression and pathway analysis of RNA-Seq data. BMC Bioinformatics 2018, 19(1):534. PMID:30567491 Full text

Usage Statistics

As of June 5, 2019, iDEP website has been visited 30,895 times by 8,252 users from 91 countries. Fully functional only in early 2018, iDEP has been used by researchers to analyze transcriptomic data from sun flowers to primates to human, as demonstrated by the 22 papers citing iDEP.



Changes

v 0.38 Gene ID conversion, remove redudancy; rlog option set to blind=TRUE.
v 0.39 Reorganized code. Updated to Bioconductor 3.5; solved problems with PREDA 9/8/17.
v 0.40 Moved libraries from the beginning to different places to save loading time.
v 0.50 Enable changing colors for heatmap. Fixed error with voom; also changed method to TMM, see here.
2/5/2018 V 0.62 : Add tree and networks to visualize overlaps between enriched gene sets, in K-means, DEG2, and pathway tags. Downloads of pathway analysis results and high-resolution figures.
2/6/2018: Fixed errors caused by gene symbol matching for unknown species. More user control of hierarchical clustering tree
2/9/2018: V 0.65 Added API access to STRINGdb website on the DEG2 tab. Supports thousands of bacterial species
2/10/2018: V 0.66 Improved API access to STRINGdb, by adding automatic species matching.
2/11/2018: V 0.67 Tested with larger dataset of 259 samples. Changed figure configurations.
2/14/2018: V 0.68 Fixed Pathview loading code. Connected Pathview to PGSEA.
2/25/2018: V 0.68 Fixed Fold-change, FDR data upload and parsing. Figure resolution using the res=150 option help improve readability of labels.
2/28/2018: V0.69 Change interactive heat maps to re-order columns
3/12/2018: V0.70 Generating R code and downloading annotation files used in analysis.
3/14/2018: V0.71 Improve R markdown file; add color to EDA plots; detect bias in sequencing depth
3/18/2018: v0.711 Fixed error caused by gene names containing characters such as ' or "
3/28/2018: v0.712 Fine tuned EDA plots
4/3/2018: V0.713 add permutations for fgsea. Expand quotes.
4/13/2018: V0.72 fixed bug caused error in R Markdown file when users choose species other than the default.
4/25/2018: V0.73 Fixed bug for GO terms tree in k-Means. Added layout button for network viz of GO terms
5/28/2018: V0.73 Enabled download of all FDR and fold-changes at the DEG1 tab. Problem with the FDR and fold-change data with only one columns.
7/28/2018: v0.80 New v0.80 Updated annotation database. Comprehensive pathway database for human. TF binding motifs for 200+ speceis. Old version made available.
7/29/2018: v0.80 Orgized UI.R according to Google R style guide.
7/30/2018: V0.80 Fixed error in downloaded up- and down-regulated gene lists. We thank Juan Xie for pointing this out.
12/2/2018: v0.81 High resolution figure download with eps format, which can be eidted with Adobe Illustrator.
3/5/2019: v0.82 Fix a bug regarding limma for identification of D.E.Gs. Up- and down-regulation are opposite in some cases.
3/29/2019: v0.85 Annotation database upgrade. Ensembl v 95. Ensembl plants v.42, and Ensembl Metazoa v.42.
5/19/2019: v0.90 Annotation database upgrade. Ensembl v 96. Ensembl plants v.43, and Ensembl Metazoa v.43. STRING-db v10


In loving memory of my parents. X.G.