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Blood, Vol. 113, Issue 12, 2826-2834, March 19, 2009
Role for MKL1 in megakaryocytic maturation
Blood Cheng et al.
113: 2826
Supplemental materials for: Cheng et al
Files in this Data Supplement:
- Table S1. List of MKL1-modulated genes in HEL cells (PDF, 18.1 KB)
- Figure S1. Murine bone marrow cells were cultured with TPO for 1 day, ckit+CD41+ megakaryocytic progenitors were sorted (day 1), and differentiated into mature megakaryocytes in vitro (day 2, day 5) (JPG, 59.4 KB)
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(A) Representative Wright-Giemsa stained cytospins for day1 and day5 samples as indicated. (B) Relative Gata1, Nf-e2, and c-kit mRNA levels were assessed by qRT-PCR and were normalized to 18S rRNA or absolute cell number.
- Figure S2. Wright-Giemsa stained cytospins of HEL/MKL1 cells in response to TPA over 4 days with or without addition of doxycycline (JPG, 86.2 KB)
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- Figure S3. Knockdown of SRF in HEL cells by siRNA (JPG, 23.6 KB)
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HEL/MKL1 cells were induced with TPA for 72 h after nucleofection with siRNA. Two validated siRNA duplexes were used to decrease SRF transcript levels in the nucleofection. Western blot is shown to determine the efficiency of SRF downregulation. Note that MKL1 overexpression levels are not affected.
- Figure S4. Microarray data validation (JPG, 47.8 KB)
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Flow cytometric confirmation that GP5 is upregulated in response to overexpression of MKL1. Representative data showing GP5 (CD42d) expression in TPA-stimulated HEL/MKL1 cells or control cells after 4 days of differentiation with or without Dox. Solid lines show GP5 expression, whereas tinted show isotype control.
- Figure S5. Representative high-power (200×) micrographs of hematoxylin and eosin staining of WT, HET, and MKL1 KO femoral bone marrow sections (JPG, 146 KB)
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Blue arrows illustrate megakaryocytes.
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