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BMPR2 regulates the self-renewal of adult HSCs
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Figure 5. BMPR-II deficient cells have reduced p38 levels and up-regulation of TJP1. (A) Western blot analysis of SMAD1/5 phosphorylation in WT and BMPR-II deficient c-kit+ cells, with and without BMP4 stimulation in vitro (n=3). ψP<0.05 compared to wild-type (WT) stimulated with BMP4 and P<0.01 compared to WT (without BMP4). (B) Western blot analysis of phospho-p38 in WT and BMPR-II-/- cells cultured over night with or without BMP4 (n=3).(C) Green fluorescent protein-positive (GFP+) cells in BM from BRE-GFP reporter mice by flow cytometry analysis (n=3). (D) Western blot analysis of total p38 in WT and BMPR-II-/- cells cultured over night with or without BMP4 (n=3). ψP<0.05 compared to WT stimulated with BMP4 and P<0.01 compared to WT (without BMP4). (E) GFP+ cells following over night in vitro culture with or without BMP4 (n=3). (F) Two separate quantitative polymerase chain reaction analyses of TJP1 expression in WT and BMPR-II deficient LT-HSC (n=3-4). Due to statis- tically detectable variability between experiments and consistent ratio based changes in expression levels, ratio paired t-test was used to compare groups. TJP1 protein levels could not be investigated as all tested commercially available antibodies yielded no results (data not shown). **P<0.01; ***P<0.001.
haematologica | 2021; 106(8)
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