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. 2019 Nov 8;14(11):e0224900. doi: 10.1371/journal.pone.0224900

Fig 2. Immunosuppressors abolish UM171 activity in HSPC.

Fig 2

A: CD34+ cord blood cells engineered to express shNT (GFP) or shNR3C1 (shGR) were cultured for 7 days with UM171 (35nM) in presence or absence of Dex (100nM). UM171-mediated expansion of HSC (CD34+CD45RA-EPCR+) subset from transduced (GFP+) cells were evaluated by flow cytometry. B: CD34+ cord blood cells were cultured for 7 days in presence of vehicle or Dex only (100nM). Representative FACS profiles show a reduction of HSCs (CD34+EPCR+) and progenitors (CD34+ and CD34+CD45RA+) in presence of Dex. C: CD34+ cord blood cells were exposed to DMSO, UM171 (35nM), Dex (100nM) or UM171 and Dex for 7 days and co-stained with CD34, EPCR and CD90 antibodies. Total cell counts and counts of HSC enriched subsets are presented. Data show mean ±SEM of 2 independent experiments performed in triplicate. D: Day 7 cultures were transplanted in immunocompromised NSG mice (outcome of 2000 day 0 cells). Human CD45 and CD34 engraftment were assessed at 26 weeks post-transplantation. E: CD34+ cord blood cells were cultures for 5 days in presence of DMSO, UM171 (35nM), NFKB inhibitor (EVP4593, 100nM) and UM171 + EVP4593. FACS profile (left panel) and absolute counts (right panel) of CD34+EPCR+, CD34+EPCR+CD90+ and CD34+CD86+ cells are shown. Data show mean ±SEM of 2 independent experiments performed in triplicate.