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Hypoxia-inducible regulation of placental BOK expression by Daochun Luo, Isabella Caniggia, and Martin Post Biochem. J. Volume 461(3):391-402 August 1,

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Presentation on theme: "Hypoxia-inducible regulation of placental BOK expression by Daochun Luo, Isabella Caniggia, and Martin Post Biochem. J. Volume 461(3):391-402 August 1,"— Presentation transcript:

1 Hypoxia-inducible regulation of placental BOK expression by Daochun Luo, Isabella Caniggia, and Martin Post Biochem. J. Volume 461(3):391-402 August 1, 2014 ©2014 by Portland Press Ltd

2 Analysis of the human BOK genomic structure (A) Schematic illustration of the human BOK promoter. Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

3 EMSA Binding of hypoxia-inducible transcription factors to the BOK-HRE was determined by gel-shift assay. Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

4 ChIP assays JEG3 cells were exposed to 21% (lanes 1–5) or 2% (lanes 6–10) O2. Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

5 Effect of of HIF proteins on BOK-HRE in the context of a heterologous promoter (A) Schematic illustration of BOK-HRE reporter constructs: control vector SV40-SEAP2, 1.0 kb BOK-SEAP2, BOK-HRE(+)-SV40-SEAP2, BOK-HRE(−)-SV40-SEAP2, mutated BOK-HRE-SV40-SEAP2 a... Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

6 Effect of hypoxia on the activity of the native BOK promoter (A) Schematic illustration of promoterless pSEAP2-basic, 1.0 kb BOK pSEAP2 and its negative control, 0.8 kb BOK-pSEAP2 missing the BOK-HRE. Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

7 Effect of hypoxia on the antisense-BOK promoter (A) Schematic illustration of promoterless pSEAP2-basic, 1.7 kb antisense-BOK pSEAP2 and its positive control, 1.0 kb BOK pSEAP2. Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

8 Knockdown of the HIF system suppresses the BOK-HRE response to hypoxia HEK-293T (A) or JEG-3 (B) cells transfected with both expression plasmids (pSEAP2-Basic, 0.8 kb-BOK- pSEAP2, 1.0 kb-BOK-pSEAP2 or 1.7 kb-Antisense-BOK-pSEAP2) and siRNAs targeting either... Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

9 Expression of antisense-BOK RNA (A) JEG3 cells without treatment of 2% O2 expressed antisense-BOK (400 bp PCR product; lanes 1 and 2). Daochun Luo et al. Biochem. J. 2014;461:391-402 ©2014 by Portland Press Ltd

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