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The concordance between RNA-seq and microarray data depends on chemical treatment and transcript abundance


The concordance of RNA-sequencing (RNA-seq) with microarrays for genome-wide analysis of differential gene expression has not been rigorously assessed using a range of chemical treatment conditions. Here we use a comprehensive study design to generate Illumina RNA-seq and Affymetrix microarray data from the same liver samples of rats exposed in triplicate to varying degrees of perturbation by 27 chemicals representing multiple modes of action (MOAs). The cross-platform concordance in terms of differentially expressed genes (DEGs) or enriched pathways is linearly correlated with treatment effect size (R2≈0.8). Furthermore, the concordance is also affected by transcript abundance and biological complexity of the MOA. RNA-seq outperforms microarray (93% versus 75%) in DEG verification as assessed by quantitative PCR, with the gain mainly due to its improved accuracy for low-abundance transcripts. Nonetheless, classifiers to predict MOAs perform similarly when developed using data from either platform. Therefore, the endpoint studied and its biological complexity, transcript abundance and the genomic application are important factors in transcriptomic research and for clinical and regulatory decision making.

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Figure 1: Overview of study design.
Figure 2: Concordance between RNA-seq and microarray.
Figure 3: Transcript abundance–dependent concordance between RNA-seq and microarray.
Figure 4: Concordance of RNA-seq and microarray data with qPCR data.
Figure 5: Cross-platform comparisons of prediction results between two platforms.
Figure 6: Systemic trends of differentially expressed RNA elements.

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Gene Expression Omnibus

Sequence Read Archive


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We thank M. Arana and D. Mendrick for their critical review of the manuscript. This research was supported, in part, by the Intramural Research Program of the National Institutes of Health (NIH), National Institute of Environmental Health Sciences (NIEHS) (ES102345-04 and ES023026) and National Library of Medicine. P.P.Ł. and D.P.K. acknowledge support by the Vienna Scientific Cluster (VSC), the Vienna Science and Technology Fund (WWTF), Baxter AG, Austrian Research Centres (ARC) Seibersdorf and the Austrian Centre of Biopharmaceutical Technology (ACBT).

Author information

Authors and Affiliations



W.T. coordinated the consortium study and manuscript preparation. W.T., S.S.A. and C.W. designed the study. C.W. conducted sequencing and qPCR experiments. S.S.A. provided rat tissue samples, gene expression data and contributed to the data analysis. P.R.B. was involved heavily in manuscript preparation and data analysis. B.G. and J.X. conducted the majority of data analysis and prepared various figures and supplementary materials. J.T.M. and D.T.M. constructed the mapping table between microarray and RNA-seq along with other data analysis and interpretation. All the co-authors contributed to various components of the study, including data analysis and preparation of text, figures, tables and supplementary materials.

Corresponding authors

Correspondence to Pierre R Bushel, Scott S Auerbach or Weida Tong.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Tables 1, 2, 4–9, 12, 13 and Supplementary Notes 1–6 (PDF 3022 kb)

Supplementary Table 3

RNA-seq data and mapping status summary based on data analysis pipeline P1 (XLSX 31 kb)

Supplementary Table 10

List of transcripts with shortened 3' UTRs detected from the samples treated by chemicals PHE and PIR (XLSX 156 kb)

Supplementary Table 11

List of differentially spliced isoforms detected in samples treated by chemicals PHE and PIR (XLSX 205 kb)

Supplementary Table 14

Master table for mapping Affymetrix probesets to RNA-seq gene annotations (XLS 8113 kb)

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Wang, C., Gong, B., Bushel, P. et al. The concordance between RNA-seq and microarray data depends on chemical treatment and transcript abundance. Nat Biotechnol 32, 926–932 (2014).

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