1. Supplementary Table 1: Microarray datasets
Supplementary Table 1: Microarray datasets. Fourteen human and one mouse CRC microarray data sets analyzed in this study from the Gene Expression Omnibus (GEO) and the ArrayExpress databases. The GEO or ArrayExpress database identifier (ID), species, microarray platform, number of individual samples run on the array (sample#), associated publication (Author), Institute of origin and the PubMed identifier for the associated publication (PMID) are provided.

2. Supplementary Table 2. Probes and primer details for the mouse putative NFAT target genes analyzed in MC38Met cells.

3. Supplementary Table 3. Probes and primer details for the RT-Ready plates used for putative NFAT target genes analyzed in human CRC patients RNA.

4. Thermoscientific Identification Target sequence
siRNA J , NFATC1
GAAUCGAGAUCACCUCCUA
siRNA J , NFATC1
CCACAGGCCUCGUAUCAGU
siRNA J , NFATC1
UAUACCAGCUCUGCCAUUG
siRNA J , NFATC1
ACGGUUACUUGGAGAAUGA
Supplementary Table 4. ON-TARGETplus SMARTpool siRNA L targeting Mouse NFATc1. ON-TARGETplus Non-Targeting pool (Cat# D )

5. Gene Symbol Full name Functional category
ANGPTL2*#
Angiopoeitin-like protein
Matrix associated protein
ANTXR1
Anthrax toxin receptor
Actin reorganization,
ASPN*#
Asporin
Bone mineralization, morphogen
BNC2*
Basonuclin
Nuclear factor
COL12A1#
Collagen 12A1
Matrix protein
COL3A1*#
Collagen 3A1
FAP*#
Fibroblast Activation Protein
GREM1#
Gremlin
Morphogen
HOPX*
HOP homeobox
Transcription factor
INHBA
Inhibin A
MGP*#
Matrix Gla Protein
MRC2*#
Mannose receptor
Endocytosis
MXRA8#
Matrix remodeling associated protein
Matrix remodeling
PRRX1*
Paired Wingless homeobox
PTRF*#
Polymerase 1 and transcript release factor
RCN3*
Reticulocalbin
ER protein, Ca2+ binding
SULF1#
Sulfatase I
Matrix remodeling enzyme
THBS3*
Thrombospondin
TWIST1*#
Twist
Morphogenesis
VCAN#
Versican
WISP1#
Wnt-inducible signaling protein
Cell adhesion/growth
Supplementary Table 5: Twenty-one putative NFAT transcriptional target genes. Gene symbols are paired with the full name of each gene and an assigned functional category is given for each. # Genes significantly down with siNFATc1 treatment, * significantly down with FK506 treatment, *# significantly down with both siNFATc1 and FK506 treatment.

6. moderated-t statistic
Gene
MC38Par-1
MC38Par-2
MC38Par-3
MC38Met-1
MC38Met-2
MC38Met-3
logfold change
Fold change
moderated-t statistic
p value
FDR
NFATC1
6.8559
7.0236
6.4217
8.2539
8.3239
8.4554
1.5773
2.9842
2.40E-06
0.0001
NFATC2
3.3507
3.4645
3.1843
3.3224
2.8869
3.7502
0.9459
0.9652
NFATC3
4.0747
4.4118
3.9329
4.2507
4.2948
4.0186
0.0482
1.0340
0.3628
0.7253
0.8141
NFATC4
5.9712
5.6826
6.135
6.6945
6.2729
6.4402
0.5396
1.4536
3.8193
0.0043
0.0176
Supplementary Table 6: Summary statistics of microarray-based differential mRNA expression of NFAT family of transcription factors in three biological replicates of MC38Par and MC38Met cell lines. Statistical significance for differential expression only for NFATc1 (bolded and highlighted in red) and NFATc4 (bolded) was achieved in this analysis.

7.  8-gene signature
16-gene signature (Ref 15)
29-gene signature (Ref 25)
149-gene signature (Ref 26)
52-gene signature (Ref 27)
449-gene signature (Ref 24)
MGP
COL3A1
PTRF
ANGPTL2
FAP
TWIST1
ASPN
Supplementary Table 7: Overlapping genes between 8-gene signature and previously published colon cancer specific prognostic signatures. Overlapping genes in referenced published gene signatures are highlighted alongside the NFAT-directed 8-gene signature described for metastasis in this model.

8. Supplementary Table 8. Summary statistics for in vivo work using isogenic MC38 cell lines in C57BL6/J mice.