SUPPLEMENTARY DATA. Table S1 Clinical features of new SS/MF patients enrolled in SNP6.0 and 10K analysis. % of TCRVß+ within CD3+CD4+

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1 SUPPLEMENTARY DATA Table S1 Clinical features of new SS/MF patients enrolled in SNP6.0 and 10K analysis Sample ID % of TCRVß+ Disease Sex/Age TCRVß Family CD4/CD8 Sample material within CD3+CD4+ 10K SNPs SNP6.0 Time of Disease (mo) Status SS 40 SS M/ % ^1,97 CD3+sorted cells y n 1 AD SS 44 SS M/79 null 50% ^3,57 CD4+sorted cells y n 4 DD SS 46 SS M/ % ^2,11 CD4+sorted cells y n 94 DD SS 53 SS M/ % ^12,50 CD4+sorted cells n y 4 DD SS 55 SS F/ % ^40,50 CD4+sorted cells y n 11 DD SS 57 SS M/ % ^6,42 CD4+sorted cells n y 4 AD SS 58 SS M/78 null 80% ^1,50 CD4+sorted cells y n 18 DD SS 60 SS M/75 null 81% ^6,3 CD4+sorted cells n y 1 DD SS 61 SS M/ % ^3,71 CD4+sorted cells y n 1 DD SS 62 SS M/80 null 90% ^47,1 CD4+sorted cells y n 13 AD SS 63 SS M/69 null 68% ^11,33 CD4+sorted cells n y 9 AD SS 64 SS M/ % ^3,9 CD4+sorted cells n y 13 AD SS 65 SS M/ % ^14,9 CD4+sorted cells n y 4 AD SS 66 SS F/ % ^5,40 CD4+sorted cells y n 13 AD SS 67 SS F % ^54,84 CD4+sorted cells n y 48 AD SS 69 SS F/61 null 94% ^45,8 CD4+sorted cells n y 17 AD SS 70 SS F/ % ^3,22 CD4+sorted cells n y 23 AD Percentage of dominant TCRVß + circulating neoplastic cells detected by fluorescence-activated cell sorting (FACS); Months between disease diagnosis and time of analysis; In this case the TCR-Vß panel was unable to identify the neoplastic clone in >95% CD4+ T cells, in the presence of a high percentage cells. (>97%) of CD60pos CD49dneg CD26neg CD7neg. This feature could be considered as an indirect proof of clonality [Scala E, Narducci MG, Amerio P et al. J. Invest. Dermatol. 119(1), (02)]. DD died of disease; AD alive with disease.

2 Table S2. Primer sequences used for genomic and mrna qrt_pcr and promoter hypermethylation analysis of PTEN and PTENP1. Gene Primers sequences Amplicon size PTEN forward primer 5 cataacgatggctgtggttg bp reverse primer 5 cccccactttagtgcacagt 3 SDC4 forward primer 5 cagggtctgggagccaagt bp reverse primer 5 gcacagtgctggacattgaca 3 COMT forward primer 5 gctactggctgacaacgtga 3 1 bp reverse primer 5 tccaccacctccctgtattc 3 PTEN-III-Methylated* forward primer 5 ggtttcggaggtcgtcggc bp reverse primer 5 caaccgaataataactactacgacg 3 PTEN-III-Unmethylated * forward primer 5 tgggttttggaggttgttggt bp reverse primer 5 acttaactctaaaccacaacca 3 PTEN forward primer 5 cataacgatggctgtggttg bp reverse primer 5 cccccactttagtgcacagt 3 PTENP1 forward primer 5 cacatttgccagaatgatga 3 82 bp reverse primer 5 tcgtcaatgtgtgaggttcc 3 β-actin forward primer 5 gatgagattggcatggcttt bp reverse primer 5 gtcaccttcaccgttccagt 3 Primers were used for genomic qrt_pcr, * for promoter hypermethylation analysis and for mrna qrt_pcr

3 Fold change of endogenous mirs and pmirs post trasfection (LogN) Figure S1. qrt_pcr analysis of mir-21, mir-106b, mir-214 and mir-486 expression evaluated in H9 cell line at basal level (grey bars) and 48 hours post transfection (black bars) using purified CD4+ healthy lymphocytes as control.. H9 Cell Line mir-21 pmir-21 (48h) mir-106b pmir-106b (48h) mir-214 pmir-214 (48h) mir-486 pmir-486 (48h)

4 Figure S2. qrt-pcr analysis for PTEN and PTENP1 expression evaluated in H9 cells 48 hours post transfection of pre-mirs using transfected FAM scramble oligos cells as control (FC=1).

5 Figure S3 IHC for pakt (DAKO) performed in psoriasis IHC for pakt (DAKO) performed in healthy skin

6 Supplementary Material & Methods Library preparation, sequencing data analysis and variants identification Libraries were prepared starting from 10 ng of genomic DNA using Ampliseq custom oligos, 2x pools (Life Technologies, cat ) and Ion Ampliseq Library kit v.2 (Life Technologies, cat ), following company s indications. Barcoded adapters containing Ion Torrent specific motifs, from Ion Xpress Barcode Adapters 1-16 and Kits (Life Technologies, cat ) were added to each sample during library preparation. Final libraries were quantified using Bioanalyzer (Agilent) with High Sensitivity DNA kit (Agilent, cat ), diluted and pooled together in the same amount. Twenty ul of 16 pm pool of all libraries were mixed to Ion Sphere Particles (Life Technologies) and clonally amplified in an emulsion PCR, performed in accordance to Ion OneTouch 0 Template kit v.2 DL protocol (Life Technologies, cat ) and using the Ion OneTouch instrument (Life Technologies, cat ). Enrichment-System (from Ion OneTouch System, Life Technologies) and Dynabeads MyOne Streptavidin C1 Magnetic Beads (Life Technologies, cat ) were used to enrich template-positive Ion Sphere Particles. Enriched sample was loaded in an Ion 316 chip (Life Technologies, cat ) and sequenced using Ion Torrent PGM, following the instructions of Ion PGM 0 Sequencing kit protocol (Life Technologies, cat ). Sequencing data analysis was conducted using Torrent Suite from Life Technologies: briefly low quality reads were removed, adapters sequences were trimmed and alignment against a reference genome (hg19) was performed using the Torrent Mapping Alignment Program. Variant Caller plugin (from Torrent Suite) was used to identify significant variations to reference genome in all samples.

7 Primer pairs used for PTEN sequencing data analysis. Amplicon ID Fwd Primer bp Rev Primer bp Amplicon size PTEN_seq1 GGCACTGTTGTTTCACAAGATGATG 25 AGTCTCCTGCATAAATTTCAAATGTGGT PTEN_seq2 TGGCTACGACCCAGTTACC 19 GCCACTGAACATTGGAATAGTTTCAAA PTEN_seq3 AGCCATTTCCATCCTGCAGAAG 22 TCCCACGTTCTAAGAGAGTGACA PTEN_seq4 GAAGATCTTGACCAATGGCTAAGTGA 26 TCTCAGATCCAGGAAGAGGAAAG PTEN_seq5 ACCTGTTAAGTTTGTATGCAACATTTC 27 GAATTGCTGCAACATGATTGTCATCT PTEN_seq6 AGATGAGTCATATTTGTGGGTTTTCA 26 GCTGATCTTCATCAAAAGGTTCATTCTC 28 9 PTEN_seq7 TGACACCACTGACTCTGATCCA 22 ATTGGTCAGGAAAAGAGAATTGTTCC PTEN_seq8 TTAACCATGCAGATCCTCAGTTTGT 25 CTGTCCTTATTTTGGATATTTCTCCCAA PTEN_seq9 GATTTGCTTGAGATCAAGATTGCAGAT 27 TTCACCTTTAGCTGGCAGACC PTEN_seq10 CCTCAGAAAAAGTAGAAAATGGAAGTCT 28 CATCACATACATACAAGTCAACAACCC PTEN_seq11 TGCAAATGTTTAACATAGGTGACAGA 26 GCAAATGCTATCGATTTCTTGATCACA PTEN_seq12 CATTGACCACCTTTTATTACTCCAGCTA 28 TGTTCCTGTATACGCCTTCAAGTC 24 0 PTEN_seq13 TTGCTATGGGATTTCCTGCAGAAA 24 CCATTAGGTACGGTAAGCCAAAAA PTEN_seq14 AATTTCAAATGTTAGCTCATTTTTGTTAATGG 32 CCTCACTCTAACAAGCAGATAACTTTCA PTEN_seq15 AACTTTTCTTTTAGTTGTGCTGAAAGACATT 31 TCACTCGATAATCTGGATGACTCATT PTEN_seq16 ACCGCCAAATTTAATTGCAG CGATAATCTGGATGACTCATTATTGT PTEN-miR106b-1 CCACAGGGTTTTGACACTTG CACATCAGTCTGTCTCCACTTTTT PTEN-miR106b-2 TTCTCAATTGTAACGACTTCTCCA 24 AGCACATGAAGCATCCACAG 146 PTEN-miR106b-3 CTGTGGATGCTTCATGTGCT GAACGGCTGACAGCTATTGA 159 PTEN-miR106b-4 CCTACATGTGCTTTATTGATTTGC 24 TGAAGCCCTAGTCCCAACTC 129