Skip to main content
. 2017 Feb 27;7(5):1100–1113. doi: 10.7150/thno.18198

Figure 7.

Figure 7

AHNAK2 knockdown impairs hypoxia-induced EMT and cancer stem cell-like features. (A) Immunoblot of CAKI-1 cells after exposed to hypoxia (1% O2) for 0, 6, 12, 24, or 48 hours for the EMT markers including E-cadherin, ZO-1, N-cadherin, Vimentin, beta-catenin, and stemness marker Nanog, an beta-actin antibody was used as loading control. (B) CAKI-1 cells that express HA-vector, HA-HIF1α vector or HA-HIF1α and AHNAK2 shRNA vectors were exposed to normoxia (21% O2) or hypoxia (1% O2) for 24 hours, and then were examined for morphology (Scale bar = 100μm). (C) The relative expression of HIF1α, AHNAK2, ZO-1, Vementin, Nanog and OCT-4 were assayed in cells as (B) by quantitative RT-PCR. * p < 0.05, ** p < 0.01, *** p < 0.001. (D) Extracts of these cells were immunoblotted for HIF1α, Nanog, and the EMT markers ZO-1, Vimentin, beta-catenin. (E) Representative images of migration assays in CAKI-1 cells stably expressing HA-vector, HA-HIF1α vector or HA-HIF1α and AHNAK2 shRNA vectors, and exposed to normoxia (21% O2) or hypoxia (1% O2) for 24 hours (scale bar = 100 μm). (F) Quantification of the migration assays of (E). * p < 0.05, ** p < 0.01, *** p < 0.001.