Page 180 - 2022_02-Haematologica-web

M.H. Lundberg Slingsby et al.
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Figure 1. The effect of antisense oligonucleotides on proplatelet production by megakaryocytes derived from human cord blood and mouse fetal liver. Human and mouse megakaryocytes (MK) were treated with vehicle (HEPES, 10 mM), puromycin (100 nM, a known inhibitor of proplatelet production), or 5 mM of the 2’MOE anti- sense oligonucleotides (ASO) 104838 or 501861 or the CpG ASO 120704 or ODN 2395 for 24 h using the IncuCyte Live Cell Analysis System and analyzed with Ilastik and Cell Profiler. (A) Proplatelet count: individual count of proplatelet protrusions that emanate from the human cord blood-derived MK body, over the course of the 24 h incubation. (B) Percent of proplatelet-producing mouse fetal liver-derived MK treated with the above treatments as well as the 2’MOE ASO 487660. *P<0.05 by two- way analysis of variance with Bonferroni post-test (n=4 repeat runs). (C) Representative brightfield images at 20x magnification, of proplatelet-producing mouse fetal liver-derived MK at 0 h, 8 h, 16 h and 24 h treated with vehicle (HEPES), the 2’MOE ASO 104838 or the CpG ASO ODN 2395. Scale bars indicate 50 mm.
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haematologica | 2022; 107(2)