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. Author manuscript; available in PMC: 2010 Sep 25.
Published in final edited form as: Mol Cell. 2009 Sep 24;35(6):856–867. doi: 10.1016/j.molcel.2009.09.006

Figure 2.

Figure 2

Mir-210 is broadly expressed and is a HIF1α-specific gene. A) Up-regulation of mir-210 by hypoxia is detected in all six different tumor types examined. Each cell line was split into two identical plates 24 hours prior to treatment. One was exposed to 2% O2 for 24 hours while the other was kept under normoxia and then, RNAs were harvested at the same time and used for Northern blotting. RCC4/VHL and 786/VHL are RCC4 and 786-O cell lines with stably reconstituted wild-type VHL gene. mir-210 is well induced by hypoxia in almost all cell lines examined, with highest induction in HSC3, MCF10A, and MCF7 cells. However, up-regulation by hypoxia is barely seen in MDA231, SQB20, and 786-O cells. U6 RNA was used as a loading control; B) Western blot of HIF1α and HIF2α in MDA-MB-231 and paired RCC4, 786-O cells. No HIF1α could be detected in 786-O and MDA-231 cells under 2% O2 for 24 hr. However, HIF2α could be readily detected in all cell lines. C) mir-210 expression was assayed by qPCR in RNAs harvested from RCC4/VHL cells with siRNA knock-down of HIF1α, HIF2α, or a scramble control siRNA with or without exposing to 2% O2 for 24 hours (lower panel). RNA input was normalized on small nuclear RNA RNU48. Upper panel shows the hypoxia induction and siRNA knock-down of HIF1α and HIF2α in these cells.