Blood, Vol. 116, Issue 15, 2812-2821, October 14, 2010

Depletion of L3MBTL1 promotes the erythroid differentiation of human hematopoietic progenitor cells: possible role in 20q– polycythemia vera
Blood Perna et al. 116: 2812

Supplemental materials for: Perna et al

Files in this Data Supplement:

• Figure S1 (JPG, 527 KB) -
  (A) Diagram of L3MBTL1 indicating the coding and 3′UTR regions targeted by the shRNAs (sh1, sh2, sh3) (B) shRNAs sequences used (C) Sh1 and sh2 were cloned into H1P-Hygro EGFP lentiviral plasmid described previously. Sh3 was cloned into pLKO-GFP plasmid with shRNA expression driven by U6 promoter and EGFP expression by HPGK promoter.
  
  
  
  
  
  
• Figure S2. Expression of GlyA in L3MBTL1-KD CD34⁺ cells at day 3, 4, and 6 of culture with SCF 100ng/ml and Epo 6 IU/ml, compared to control vector (JPG, 500 KB) -
  
  
  
  
  
  
• Figure S3. Levels of GATA-1, FOG-1, EPOR, FKLF, NF-E2, and LMO2 mRNA expression in L3MBTL1-KD CD34⁺ cells plated in Epo-containing cultures, compared to controls (JPG, 359 KB) -
  
  
  
  
  
  
• Figure S4. Expression of Glycophorin A in L3MBTL1-KD K562 cells treated with hemin for 48 hours (JPG, 473 KB) -
  
  
  
  
  
  
• Figure S5 (JPG, 373 KB) -
  (A) Frequency of CD34⁺ CD38⁺ double positive cells in control and L3MBTL1-KD cells and the percentage of CMP, GMP, and MEP cells among the GFP positive L3MBTL1-KD or control CB cells. (B) L3MBTL1 mRNA expression levels in CMP, GMP, and MEP cells, isolated from cord blood. (C) Scheme representing the selection used to isolate the three human progenitor cell populations: CMP (Lin⁻, CD34⁺, CD38⁺, IL3Rα⁺, CD45RA⁻), GMP (Lin⁻, CD34⁺, CD38⁺, IL3Rα⁺, CD45RA⁺), MEP (Lin⁻, CD34⁺, CD38⁺, IL3Rα⁻, CD45RA⁻).¹⁶