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. 2020 Sep 11;9(9):81. doi: 10.1038/s41389-020-00265-z

Fig. 5. HIFs are required for hypoxia-induced SOX2 expression.

Fig. 5

a, b Validation of the siRNAs targeting HIF-1α and HIF-2α by qRT-PCR (a) and western blot (b). c Images of ECSCisk morphology with low-HIF expression subjected to 2 or 21% O2. d, e qRT-PCR analysis of stemness genes (ALDH1, CD133, OCT4, SOX2 and NANOG) (d) and lineage-specific genes (E-cadherin, Cytokeratin8, α-SMA, Vimentin and N-cadherin) (e) in ECSCisk under different conditions, as shown in the panels. f The number of ECSCisk colonies was calculated after transfection with a HIF siRNA. g The number of ECSCisk colonies was calculated after transfection with a HIF shRNA. h, i Confocal images of SOX2 and NANOG in ECSCisk subjected to various treatments. j SOX2, NANOG and E-cadherin protein levels were detected in ECSCisk transfected with control and HIF siRNAs. k FACS analysis was used to determine the percentages of CD133-positive ECSCisk under hypoxic or normoxic conditions. Scale bar = 25 μm. *P < 0.05, **/##P < 0.01, and ***/### P < 0.001.