Figure 9.

Overexpression of EZR reverses the functional effects of ANRIL knockdown in HCAECs. A–H, four different groups of HCAECs co-transfected with pcDNA3.1 vector + siNC, pcDNA3.1 vector + siANRIL, pcDNA3.1-EZR + siNC, and pcDNA3.1-EZR + siANRIL, respectively, were used for further analysis (n = 5). To maximize accuracy and consistency in data, experiments in HCAECs with CLIP1 (Fig. 7), EZR (Fig. 9), and LYVE1 (Fig. 11) overexpression plasmids were performed simultaneously, sharing the same pcDNA3.1 vector + siNC and pcDNA3.1 vector + siANRIL cotransfected controls. Data were normalized to the value of each pcDNA3.1 vector + siNC group, which was defined as 1.0. *, p < 0.05; **, p < 0.01. Only statistically significant differences are marked with asterisks. A, protein expression of Ezrin was measured in the four different HCAEC groups using Western blot analysis. GAPDH was used as a loading control. B, Western blotting images in A were quantified and plotted. C, relative overexpression of EZR was determined in the four different HCAEC groups through qRT-PCR analysis, and the results were plotted. D, relative knockdown efficiency of siANRIL in the four different HCAEC groups was determined through qRT-PCR analysis, and the results were plotted. E, adhesion of THP-1 cells to the four different HCAEC groups was quantified after 1 h of co-incubation, and the results were plotted. F, transmigration of THP-1 cells across a layer of the four different HCAEC groups was quantified after 24 h of co-incubation, and the results were plotted. G, raw images of the four different HCAEC groups at the 0- and 24-h time points during their migration. For control groups, images of the vector + siNC and vector + siANRIL cotransfected controls in Fig. 7G are reused. H, the area of the changes in the healed wound shown in G was quantified and plotted. Error bars, S.D.