Fig. 3

Effect of HIF-1α on AJC marker expression in OSCC cell lines. a Western blot of protein levels (left) and quantification (right) of AJC markers in OSCC cell lines treated with siRNA-HIF-1α or control siRNA. The experiments were performed in triplicates. b Localization of Par3 (Green) and TJP1 (Red) in SCC-25 cell line treated with siRNA-HIF-1α or control siRNA as analyzed by IF staining. White boxes are magnified images of the dashed line boxes. Scale bars: 20 μm. The experiments were performed in triplicates. c Interaction between Par3 and TJP1 in SCC-25 cell line treated with siRNA-HIF-1α or control siRNA as analyzed by in situ PLA. Red spots represent PLA positive signals. The experiments were performed in triplicates. d Cell proliferation ability of OSCC cells treated with siRNA-HIF-1α or control siRNA as analyzed by the EdU assay. Green colored cells are EdU-positive cells. Scale bars: 20 μm. The nuclei were stained with DAPI (blue). The experiments were performed at least thrice. e Representative images (left) and numbers (right) of migrated and invaded OSCC cells treated with siRNA-HIF-1α or control siRNA. Scale bars: 100 μm. The experiments were performed at least thrice. f Activities (left) and fold intensities (right) of MMP-9 and MMP-2 in OSCC cells treated with siRNA-HIF-1α or control siRNA as analyzed by gelatin zymography. *P < 0.05 vs. control siRNA. The experiments were performed in triplicates. Cont: control siRNA; siR-HIF1A: siRNA- HIF-1α; Hyp: hypoxia; PLA+: PLA positive signals in OSCC cells; EdU+: EdU-positive cells