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. 2015 Sep;100(9):1160–1171. doi: 10.3324/haematol.2014.120295

Figure 1.

Figure 1.

Effect of mild or severe hypoxia on proliferation, differentiation, HIF-1α, HIF-2α, CXCR4 and miR-146a expression in hematopoietic progenitor cells induced to selective monocytic differentiation. (A–C) Severe hypoxia (1% O2) impairs cell proliferation (A) and delays monocytic differentiation of CD34+ hematopoietic progenitor cells (day 0), as shown by flow cytometry analysis of specific monocytic markers CD14 (B) and CD11b (C) expression, as compared to mild hypoxia (5% O2). (D, E) Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) shows that 1% O2 up-regulates miR-146a (D) and CXCR4mRNA (E) expression in monocytic differentiating hematopoietic progenitor cells, as compared to 5% O2. (F) Flow cytometry analysis indicates that CXCR4 membrane protein levels are down-regulated by 1% O2, as compared to 5% O2 (G, I) qRT-PCR shows the differential expression of HIF-1α and HIF-2α mRNA during monocytic proliferation and differentiation of hematopoietic progenitor cells in hypoxia (mild or severe). (H, J) Western blot analysis of HIF-1α and HIF-2α nuclear protein expression during monocytic differentiation under 5% O2 and 1% O2 hypoxia. Nucleolin is shown as an internal control of nuclear proteins. (A, D–G, I) Mean ± SEM values from three independent experiments are shown. *P<0.05, **P<0.01. (B, C, H, J) One representative experiment of three is shown.