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Blood, Vol. 112, Issue 5, 1942-1950, September 1, 2008
Transcriptional repression of c-Myb and GATA-2 is involved in the biologic effects of C/EBP in p210BCR/ABL-expressing cells
Blood Soliera et al.
112: 1942
Supplemental materials for: Soliera et al
Files in this Data Supplement:
- Table S1. C/EBPα-regulated genes identified by nucleotide array (XLS, 55.5 KB)
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Normalized data from RNA of 4-HT–treated (0, 6, 9 and 12 hours) 32D-BCR/ABL cells expressing Δ (177-191) C/EBPα-ERTAM, K298E C/EBPα-ERTAM, or transduced with MigRI were subjected to ANOVA. Qualifiers had to meet the following criteria to be included in the analyses: be equal or greater than 43 particles per million ( ppm), be present in at least one sample and show a fold change of more than threefold relative to the 0 time-point. This identified 119 qualifiers, (28 repressed and 91 induced) which were regulated by Δ (177–191) C/EBPα in a DNA binding dependent manner.
- Figure S1. Effect of ectopic c-Myb expression on C/EBPα-induced Gr-1 expression in C/EBPα-ER-32D-BCR/ABL cells (JPG, 91.7 KB)
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(A) Gr-1 levels detected by flow cytometry; (B) Expression of endogenous and Δ (358–452) c-Myb in 4-HT–treated parental and c-Myb-expressing C/EBPα-ER 32D-BCR/ABL cells.
- Figure S2. Effect of full-length c-Myb expression on C/EBPα-induced differentiation of K562 cells (JPG, 92.1 KB)
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(A) CD11b and CD15 levels (mean plus SD of two experiments); (B) morphology; and (C) ectopic protein levels in 4-HT–treated K562 cells retrovirally-transduced C/EBPα-ER and HA-tagged c-Myb.
- Figure S3. Differentiation marker and gene expression analysis in 4-HT or c-Myb siRNA-treated C/EBPα-ER/ K562 cells (JPG, 119 KB)
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CD11b,CD15 or CD11b/CD15 (A),G-CSFR (B), PU.1 (C) and M-CSFR (D) expression in C/EBPα-ER/K562 cells treated with 4-HT and control or c-Myb siRNAs. Representative of three different experiments.
- Figure S4. Effect of GATA-2 silencing on C/EBPa-induced differentiation and proliferation inhibition in K562 cells (JPG, 110 KB)
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GATA-2 and PU.1 (A) and CD11b and CD15 (B) levels in C/EBPa-ER/K562 cells treated with 4-HT and a control or GATA-2 siRNAs ;cell counts (C) of siRNA-, 4-HT–, or siRNA-, and 4-HT–treated K562 cells. Representative of three different experiments.
- Figure S5. Effect of c-Myb silencing in 4-HT-treated C/EBPα-ER retrovirally-transduced normal (upper panel) and CML-BC (lower panel) CD34+ cells (JPG, 102 KB)
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Histogram shows CD11b expression; Western blot shows c-Myb levels in lysate of c-Myb–silenced CD34+ cells. Western blot was not performed in c-Myb–silenced CML-BC cells because cell extract was insufficient.
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