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. 2018 Sep 12;8:324. doi: 10.3389/fcimb.2018.00324

Figure 3.

Figure 3

The high-virulence H. parasuis infection induced β-catenin to translocate into nuclear in PK-15 and NPTr cells. (A,B) PK-15 cells (A) or NPTr cells (B) were seeded at a density of 5 × 104 on coverslips in 24-well plates. After culturing overnight, cells were infected or uninfected with SH0165 or HN0001 for 12 h. LiCl was used as the positive control. Cells were stained with anti-β-catenin antibody and then visualized with AlexaFluor 488-conjugated anti-rabbit IgG. Nuclei were stained with DAPI. (C,D) Protein fractions of membrane, cytoplasm, and nuclei were isolated and enriched from PK-15 cells (C) or NPTr cells (D) at 24 h after being stimulated or unstimulated with H. parasuis (108 CFU/mL) or LiCl (20 mM), and subsequently analyzed for β-catenin expression by Western blot. Caveolin-1, GAPDH, and PCNA were used as loading controls for the membrane, cytoplasmic, and nuclear fractions, respectively. Relative β-catenin levels were calculated using Image J software and normalized to β-actin. Values are presented as the ratio of each sample to the untreated control (set to 1.0). Representative results of three independent experiments are shown as the mean +/− SD (n = 3). *p < 0.05; **p < 0.01 compared with the uninfected control.