Supplementary Data

Supplementary Table 1. Demographicandclinicalcharacteristics of17patients with chronic myeloid leukemia (CML) in chronic phase.

Patient Number / Sex / Age / Diagnosis / % of BCR-ABL/ABL
1 / Male / 60 / CML / 93.5
2 / Female / 44 / CML / 132.5
3 / Male / 39 / CML / 99.12
4 / Male / 30 / CML / 132
5 / Female / 54 / CML / 43.6
6 / Male / 48 / CML / 27.38
7 / Male / 53 / CML / 18.4
8 / Male / 40 / CML / 70.38
9 / Male / 14 / CML / 82.69
10 / Male / 32 / CML / 45.73
11 / Male / 21 / CML / 302.14
12 / Male / 65 / CML / 199.12
13 / Male / 32 / CML / 171.9
14 / Male / 45 / CML / 80.5
15 / Male / 60 / CML / 102
16 / Male / 42 / CML / 165.2
17 / Male / 17 / CML / 35.54
18 / Female / 28 / CML / 15

Supplementary Table2. Clinical characteristics of 5 CML patients(chronic phase) treated with Imatinib.

Number / Sex / Age / Diagnosis / % of BCR-ABL/ABL (before Imatinib treatment) / % of BCR-ABL/ABL (after Imatinib treatment)
Patient A / Male / 37 / CML / 290 / 3.76
Patient B / Male / 12 / CML / 100 / 2.4
Patient C / Male / 29 / CML / 112 / 1.63
Patient D / Male / 60 / CML / 102 / 2.55
Patient E / Male / 44 / CML / 24.6 / 0.53

Supplementary Table 3. DNA and RNA sequences used in this study.

Name / Sense / antsense
RNA / control RNA* / UUCUCCGAACGUGUCACGUTT / ACGUGACACGUUCGGAGAATT
miR-138 / AGCUGGUGUUGUGAAUCAGGCCG / GCCUGAUUCACAACACCAGGUAU
control inhibitor** / UCUACUCUUUCUAGGAGGUUGUGA
miRNA-138 inhibitor*** / CGGCCUGAUUCACAACACCAG
siGATA11 / GGUACUCAGUGCACCAACUTT / AGUUGGUGCACUGAGUACCTT
siBCR-ABL2 / GCAGAGUUCAAAAGCCCUUTT / AAGGGCUUUUGAACUCUGCTT
siCCND33 / GAUGCUGGCUUACUGGAUGTT / CAUCCAGUAAGCCAGCAUCTT
Name / Forword primer / Reverse primer
qRT-PCR / pri-miR-138-1 / AGCAGCACAAAGGCATCTCTA / GGAGCTGGAATGCTGCCTGTA
pri-miR-138-2 / GCCACAGCCCAGAGGAGGAAG / CTGGAGAGGATGGGTATGATG
GATA1 / TGCCAGTGGGGATCCCGTGT / CCAGCTGGTCCTTCGGCTGC
FOG1 / CAACTGCGAGCGGCACCTCA / AGGCTCCGCTCTGGCCTCTC
EKLF / GCCATCCAAGCGAGGCCGA / GTCCGAGCGCGCGAATCTC
EPOR / CCAGCTCCCAGCTCTTGCGT / GAGGCTCAGCGGCTGGGATAA
Gfi-1B / CAGCCCTGTCCTTAGCACTC / GCTTGTAGAATGGGGGTGAG
p45 / AAGGTACCCGCTGACAGAGA / TGCTGGAAAATGTCACGGTA
BCR-ABL / gtgtgaaactccagactgtc / caaaatcatacagtgcaacga
BCR / CGGAGGCCAACCTGCTCACC / AGCAGCGTGGGGCCAAAGAC
ABLIa / CCAAGAAGGGGCTGTCCTCGTCC / GTTCCAACGAGCGGCTTCACTC
ABLIb / CAAGCTTGCCTGCCCTGCATT / TACTGGCCGCTGAAGGGCTTCA
CCND3 / tggatgctggaggtatgtga / gcacagtttttcgatggtca
CDK6 / tgtttcagcttctccgaggt / cgtgacgaccactgaggtta
E2F2 / ccttggaggctactgacagc / ccacaggtagtcgtcctggt
E2F3 / ggaaggcatccacctcatta / tttgacaggccttgacactg
G3PDH / TCAGTGGTGGACCTGACCTG / TGCTGTAGCCAAATTCGTTG
ChIP / Fragment1 / cataaagtgcaacttagtgct / ggatgcagggctggtgtgcag
Fragment2 / gtggtcactggttggcagtta / caatcgcacaatcagcttgta
Fragment3 / ggaagtgcccattttacactg / gtcagcaaatgctggacactg
Fragment4 / ctgccacttgtttccaaccga / gcacggcccagcacacaaaga
Vector Construction / pGL-CCND3 / GATCTGCAGctctggagtggccactaagca / GATGAATTCtacagtaggatgatggtcctg
pGL-CCND3-mut / cagaattggatacataatgcacaattccttt
tgaacgc / gcgttcaaaaggaattgtgcattatgtatc
caattctg
pGL-ABL-3`-UTR / ACGTCTAGAACAAGGGACTAGTGAGTCA / CCGGAATTCTTCTAATGTAAACACTGAT
pGL-ABL-CDSsite1 / ACGTCTAGAgaagcccttcagcggccagta / gatGAATTCagcttgtgcttcatggtgatg
pGL-ABL-CDSsite2 / ACGTCTAGAcaggtgacccgaggcacagta / gatGAATTCacctctgcactatgtcactga
pGL-ABL-CDSsite2mu / ctcaggtgcaccagggggtagactccaagg
gccccgccgagga / tcctcggcggggcccttggagtctaccccc
tggtgcacctgag
pcDNA-Flag-ABL / GATGAATTCatgttggagatctgcctgaagc / GCACTCGAGctacctctgcactatgtcactga
pcDNA-FLAG-ABLmu / ctcaggtgcaccagggggatactccaaggg
ccccgccgagga / tcctcggcggggcccttggagtatccccctgg
tgcacctgag

* The control RNA for miRNA was duplex short RNA, and did not target any endogenous gene.

**The Control inhibitor was one kind of single strand methoxyl modified oligonucleotide sequence, did not target any endogenous gene.

***The miR-138 inhibitor is the methoxyl modified antisense oligonucleotide targeting miR-138.

All miRNA, siRNA, miRNA inhibitor, and corresponding control RNA were synthetized by GenePharm company.

Supplementary Figures:

Figure S1. Flow cytometric analysis ofK562, Ku812 and bone marrow mononuclear cells transfected with 100 nM FAM-conjugated negative control RNA. The percentage of FAM-positive cells at 24 h after transfection isindicated.

Figure S2. miR-138 inhibitor and control inhibitor were transfected into K562 cells, and imatinib was added 24 h later.Cell cycle (a) and Apoptosis (b) wereanalyzed by flow cytometry as described in methods 48 h post transfection.

Figure S3.miR-138 and control RNA were transfected into K562 cells, and apoptotic cells were detected by annexin V staining 96 h later.

Figure S4. The expression ofmiR-138 was regulated by GATA1. (a) siRNA against GATA1 and control RNA were transfected into K562 cells, imatinib was added 24 h later. Total RNAs were extracted and miR-138 expression levels were detected by qRT-PCR 72 h post transfection. (b) Chromatin immunoprecipitations (ChIP) were performed of K562 cells treated with imatinib. Recovered DNA was PCR amplified with primers targeting fragments 1and 4 as indicated in Figure 6d.

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