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Fig. 6 | Cell & Bioscience

Fig. 6

From: Hypoxic metabolism in human hematopoietic stem cells

Fig. 6

Cooperative role of Pbx1, HoxA9, and Meis1 for transcriptional regulation of Hif-1α. a) Figure shows conserved consensus Meis1, Pbx1, and HoxA9 motifs found on Hif-1α gene. Note the binding motifs found next to each other in close proximity and highly conserved. b) Luciferase reporter assays demonstrate dose-dependent transcriptional activation of Hif-1α (Hif-1α-pGL2 reporter) by Pbx1 and mutation of Pbx1 bindings sites abolished Hif-1α activation (PbxMut-Hif-1α-pGL2 reporter) c) Luciferase reporter assays demonstrate dose-dependent transcriptional activation of Hif-1α (Hif-1α-pGL2 reporter) by HoxA9 and mutation of HoxA9 bindings sites abolish Hif-1α activation (HoxMut-Hif-1α-pGL2 reporter) d) Luciferase reporter assays with different combinations of Meis1, Pbx1, and HoxA9 demonstrates (additive) cooperation of Meis1, Pbx1 and HoxA9 for activation of Hif-1α (Hif-1α-pGL2 reporter) e) Schematic of Meis1, Pbx1 and HoxA9 domains f) Meis1, Pbx1 and HoxA9 protein interaction were assessed by deletion of pbx1 interaction domain at Meis1 (71-100aa HR1 and 136-180aa HR regions) and Meis1 interaction domain of HoxA9 (1-61aa). Transcriptional activation by WT vectors, and vectors with deleted domains were evaluated using Hif-1α-pGL2 construct. Luciferase measurements were calculated as firefly luciferase units versus b-gal units. g) Schematic of proposed model showing regulation of Hif-1α expression by a complex of Meis1/Pbx1/HoxA9 proteins in hematopoietic stem cells

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