Limits...
The polycomb group protein L3MBTL1 represses a SMAD5-mediated hematopoietic transcriptional program in human pluripotent stem cells.

Perna F, Vu LP, Themeli M, Kriks S, Hoya-Arias R, Khanin R, Hricik T, Mansilla-Soto J, Papapetrou EP, Levine RL, Studer L, Sadelain M, Nimer SD - Stem Cell Reports (2015)

Bottom Line: Indeed, knockdown of L3MBTL1 promotes the development of hematopoiesis and impairs neural cell fate in human pluripotent stem cells.We also found a role for L3MBTL1 in regulating SMAD5 target gene expression in mature hematopoietic cell populations, thereby affecting erythroid differentiation.Taken together, we have identified epigenetic priming of hematopoietic-specific transcriptional networks, which may assist in the development of therapeutic approaches for patients with anemia.

View Article: PubMed Central - PubMed

Affiliation: Molecular Pharmacology and Chemistry Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. Electronic address: pernaf@mskcc.org.

Show MeSH

Related in: MedlinePlus

Impaired Development of Neural Progenitors in L3MBTL1-KD Pluripotent Stem Cells(A) KD of L3MBTL1 impairs the ability to form neuronal progenitor cells and neurons. Nestin and Tuj1 expression were evaluated in EBs by IF. DAPI served as control. (Top) The first line shows multiple EBs at lesser magnification (4×); the second line shows a single EB at 10× magnification. (Bottom) The impairment in neural progenitor formation of the L3MBTL1-KD EBs. The graph on the right shows the percentage of EBs containing Tuj1+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test.(B) KD of L3MBTL1 enhances CD34 expression. (Top left) The first line shows few positive CD34+ EBs at low magnification (4×) in the controls; the second line shows a single EB at higher magnification (10×). (Bottom left) Increased CD34 staining in the L3MBTL1-KD EBs at 4× and 10× magnification is apparent. (Top right) Increased expression of CD34 marker (21%) in the L3MBTL1-KD cells compared to controls (3%) by FACS is shown. (Bottom right) The graph gives the percentage of EBs containing CD34+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test. See also Figure S2.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4400644&req=5

fig2: Impaired Development of Neural Progenitors in L3MBTL1-KD Pluripotent Stem Cells(A) KD of L3MBTL1 impairs the ability to form neuronal progenitor cells and neurons. Nestin and Tuj1 expression were evaluated in EBs by IF. DAPI served as control. (Top) The first line shows multiple EBs at lesser magnification (4×); the second line shows a single EB at 10× magnification. (Bottom) The impairment in neural progenitor formation of the L3MBTL1-KD EBs. The graph on the right shows the percentage of EBs containing Tuj1+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test.(B) KD of L3MBTL1 enhances CD34 expression. (Top left) The first line shows few positive CD34+ EBs at low magnification (4×) in the controls; the second line shows a single EB at higher magnification (10×). (Bottom left) Increased CD34 staining in the L3MBTL1-KD EBs at 4× and 10× magnification is apparent. (Top right) Increased expression of CD34 marker (21%) in the L3MBTL1-KD cells compared to controls (3%) by FACS is shown. (Bottom right) The graph gives the percentage of EBs containing CD34+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test. See also Figure S2.

Mentions: To assess the neural differentiation potential of the L3MBTL1-KD cells, we generated EBs and cultured them in BDNF-containing conditions that support the development of neural progenitor cells (Figure 2A, top). While control EBs developed into Nestin+ neural precursors and Tuj1+ differentiated neurons, L3MBTL1-KD cells showed impaired neural lineage differentiation (Figure 2A, bottom and right). The impaired generation of neural precursors is not dependent on decreased cell proliferation or increased apoptosis, as we found no significant change in the expression of Ki-67 and Annexin V (Figures S2A and S2B).


The polycomb group protein L3MBTL1 represses a SMAD5-mediated hematopoietic transcriptional program in human pluripotent stem cells.

Perna F, Vu LP, Themeli M, Kriks S, Hoya-Arias R, Khanin R, Hricik T, Mansilla-Soto J, Papapetrou EP, Levine RL, Studer L, Sadelain M, Nimer SD - Stem Cell Reports (2015)

Impaired Development of Neural Progenitors in L3MBTL1-KD Pluripotent Stem Cells(A) KD of L3MBTL1 impairs the ability to form neuronal progenitor cells and neurons. Nestin and Tuj1 expression were evaluated in EBs by IF. DAPI served as control. (Top) The first line shows multiple EBs at lesser magnification (4×); the second line shows a single EB at 10× magnification. (Bottom) The impairment in neural progenitor formation of the L3MBTL1-KD EBs. The graph on the right shows the percentage of EBs containing Tuj1+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test.(B) KD of L3MBTL1 enhances CD34 expression. (Top left) The first line shows few positive CD34+ EBs at low magnification (4×) in the controls; the second line shows a single EB at higher magnification (10×). (Bottom left) Increased CD34 staining in the L3MBTL1-KD EBs at 4× and 10× magnification is apparent. (Top right) Increased expression of CD34 marker (21%) in the L3MBTL1-KD cells compared to controls (3%) by FACS is shown. (Bottom right) The graph gives the percentage of EBs containing CD34+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test. See also Figure S2.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400644&req=5

fig2: Impaired Development of Neural Progenitors in L3MBTL1-KD Pluripotent Stem Cells(A) KD of L3MBTL1 impairs the ability to form neuronal progenitor cells and neurons. Nestin and Tuj1 expression were evaluated in EBs by IF. DAPI served as control. (Top) The first line shows multiple EBs at lesser magnification (4×); the second line shows a single EB at 10× magnification. (Bottom) The impairment in neural progenitor formation of the L3MBTL1-KD EBs. The graph on the right shows the percentage of EBs containing Tuj1+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test.(B) KD of L3MBTL1 enhances CD34 expression. (Top left) The first line shows few positive CD34+ EBs at low magnification (4×) in the controls; the second line shows a single EB at higher magnification (10×). (Bottom left) Increased CD34 staining in the L3MBTL1-KD EBs at 4× and 10× magnification is apparent. (Top right) Increased expression of CD34 marker (21%) in the L3MBTL1-KD cells compared to controls (3%) by FACS is shown. (Bottom right) The graph gives the percentage of EBs containing CD34+ cells. The data represent the mean ± SD of the three independent experiments. ∗p < 0.05 by Student’s t test. See also Figure S2.
Mentions: To assess the neural differentiation potential of the L3MBTL1-KD cells, we generated EBs and cultured them in BDNF-containing conditions that support the development of neural progenitor cells (Figure 2A, top). While control EBs developed into Nestin+ neural precursors and Tuj1+ differentiated neurons, L3MBTL1-KD cells showed impaired neural lineage differentiation (Figure 2A, bottom and right). The impaired generation of neural precursors is not dependent on decreased cell proliferation or increased apoptosis, as we found no significant change in the expression of Ki-67 and Annexin V (Figures S2A and S2B).

Bottom Line: Indeed, knockdown of L3MBTL1 promotes the development of hematopoiesis and impairs neural cell fate in human pluripotent stem cells.We also found a role for L3MBTL1 in regulating SMAD5 target gene expression in mature hematopoietic cell populations, thereby affecting erythroid differentiation.Taken together, we have identified epigenetic priming of hematopoietic-specific transcriptional networks, which may assist in the development of therapeutic approaches for patients with anemia.

View Article: PubMed Central - PubMed

Affiliation: Molecular Pharmacology and Chemistry Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. Electronic address: pernaf@mskcc.org.

Show MeSH
Related in: MedlinePlus