Supplementary Figure S1 Supplementary Figure S2

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Supplementary Figure S1 Supplementary Figure S2 10.0 MDA-MB-435 a-tubulin p-Akt S473 Ad-LacZ Ad-TIS21 7.5 Number of cells (x105) 5.0 2.5 0.0 siCont + - + - siAkt1 - + - + Ad-LacZ Ad-TIS21

Supplementary Figure S3 DCF intensity (fold) 0.0 0.2 0.4 0.6 1.2 0.8 1.0 1.4 Ad-LacZ Ad-TIS21 MCF-7 MDA-MB-468 SK-BR-3 MDA-MB-231

Supplementary Figure S4 b c Gelatin-FITC Cont DPI FITC-gelatin F-actin Merge No. of invadopodia/ cell # Invadopodia assay Invaded cells/ field (relative value) Cont DPI 0.0 0.5 1.0 2.0 * Matrigel assay 1.5

Supplementary Figure S5 c d MitoSOXTM 25um Ad-LacZ Ad-TIS21 H2O2 Green Red Ratio SHP-Mito Ad-LacZ Ad-TIS21 50 moi Ad-TIS21 100 moi 1.0 0.8 * b 0.6 Fred/Fgreen (relative intensity) MitoSOX™ 0.5 1.0 2.0 1.5 50 200 400 - - - - - - 50 200 400 Adenovirus (moi) Ad-LacZ Ad-TIS21 0.4 0.2 0.0 Ad-LacZ Ad-TIS21 H2O2

TIS21/BTG2/Pc3 mRNA expression Supplementary Figure S6 a b c # HDF-PD14 HDF-PD85 # # # TIS21/BTG2/Pc3 mRNA expression (normalized to GAPDH) (normalized to GAPDH) mRNA expression (normalized to GAPDH) mRNA expression mDia1 mDia2 mDia3 TIS21/BTG2/Pc3 mDia1 mDia2 mDia3 HDF-PD14 HDF-PD85

a b c d Supplementary Figure S7 Epidomics antibody Santa Cruz antibody Ad-LacZ Ad-TIS21 p-Akt1 (Epitomics) Nucleus Merge p-Akt1 (Santa Cruz) Nucleus Merge Ad-LacZ Ad-TIS21 c d p-Akt1 in cytosol and nucleus (%) p-Akt1 in nucleus only (%) # Epidomics antibody p-Akt1 in cytosol and nucleus (%) p-Akt1 in nucleus only (%) # Santa Cruz antibody

a b Supplementary Figure S8 Akt2 mRNA expression (Normalized to GAPDH) siCont siAkt2 # # # # n.s (mDia1/GAPDH) (mDia2 / GAPDH) (mDia3 / GAPDH) n.s n.s siCont + - + - siCont + - + - siCont + - + - siAkt2 - + - + siAkt2 - + - + siAkt2 - + - + Ad-TIS21 Ad-TIS21 Ad-TIS21 Ad-LacZ Ad-LacZ Ad-LacZ

a b Supplementary Figure S9 Class IA IB II III n.s n.s ** (normalized to GAPDH) mRNA expression ** * C2g p110a p110b p110d p110g C2a C2b Vps34 Class IA IB II III

TIS21/BTG2/Pc3 mRNA expression Supplementary Figure S10a Normal (n=41) Tumor TIS21/BTG2/Pc3 mRNA expression transformed by log2 *p=0.006 vs. normal Wilcoxon Signed Ranks Test *

TIS21/BTG2/Pc3 mRNA expression Supplementary Figure S10b Wilcoxon Signed Ranks Test *p=0.242 vs. normal Normal (n=25) Tumor TIS21/BTG2/Pc3 mRNA expression transformed by log2 LN negative tissue (n=16) *p=0.003 vs. normal LN positive tissue *

Supplementary Figure S11a Akt1 mRNA expression transformed by log2 Normal (n=41) Tumor Wilcoxon Signed Ranks Test *p=0.010 vs. normal *

Supplementary Figure S11b Akt1 mRNA expression transformed by log2 Normal (n=16) Tumor Wilcoxon Signed Ranks Test *p=0.026 *p=0.135 (n=25) LN negative tissues LN positive tissues *

Supplementary Fig.S12 (TIS21/GAPDH, x 103) ** siCont + siTIS21 - Ad-LacZ siTIS21 siCont Ad-TIS21 - + (TIS21/GAPDH, x 103) **

Supplementary Table 4 Potential binding site for transcription factors observed in the promoter region of Nox4 examined by Alibaba2.1 Transcription Factor Number Sp1 47 MyoD 1 AP-1 3 MIG1 NF-kb 4 TFIID 2 C/EBPalpha 24 REB1 C/EBPbeta Krox-20 C/EBPgamma CREBP1 GATA-1 AFP1 Myogenin ER GCN4 TCF-1 alpha USF PTF1-beta GR 5 CD1 OCT-1. 9 NF-muE1 TBP MEF-2 HNF-1 6 TEC1 PR Tra-1 Pit-1a c-Jun CREBP eps Antp HNF-3B ID3 HNF-1C NF-ATC3 MAL63 Erg-1 OCT-2.1 HOXA1 AP-2 alphaA CPE_bind AP-2 alpha Hb AP-2 WT1 NF-1 ICSBP RAP1