Figure 5.

The YAP-TEAD transcriptional complex was involved in the regulation of nicotine-induced BiP expression and tumor malignancy in OSCC. OE and SAS cells transfected with non-targeting siRNA or YAP siRNA were exposed to 1 μM nicotine for 48 h. (A,B) The expressions of phospho-YAP (Ser127), YAP, and BiP were analyzed by quantitative RT-PCR and Western blot analysis. (C) The graphs show the quantification of Western blots. Band intensities were quantified using ImageJ software. The relative protein expressions of BiP and YAP were normalized to GAPDH expression. The abilities of cell migration (D) and invasion (E) were investigated using wound-healing and Transwell invasion assays, respectively. Black solid lines on the acquired images indicate the wound borders at 0 and 18 h post-scratching (D, left panels). Quantitative results of migratory cells across the wound borders (D, right panels) and PI-stained invasive cells (E, right panels) were determined using ImageJ software. The migratory ability was calculated by the area reduction at 18 h compared to the wound area at 0 h. N, nicotine. SC, non-targeting siRNA-transfected cells. YAP-KD, YAP siRNA-transfected cells. GAPDH served as the loading control. * p < 0.05 by Student’s t-test or one-way ANOVA followed by Bonferroni’s post hoc test. Data are presented as the mean ± SEM of three independent experiments. SEM, error bars. Scale bar, 100 μm.