Fig. 5

Phosphorylation of NF-κBp65 Ser536, but not Ser276 and Ser529, was upregulated by inflammation-mediated ARRB1. a ARRB1 transfection increased the expression of p65 Ser536, but not p65 Ser276 and Ser529, in HepG2 and HepG3B cell lines. b Cells were infected with ARRB1 or control lentivirus in LO2, HepG2, HepG2.2.15, and Hep3B cell lines. Cells stably expressed ARBB1 after transfection and selection by puromycin. c, d ARRB1 transfection increased the expression of p-p65, but not p65, in LO2, HepG2 and Hep3B cell lines. e Fluorescence images showed induced nuclear p65 expression in LO2, HepG2, HepG2.2.15, and Hep3B cell lines following transfection of ARRB1 using the immunofluorescence assay. f p65 was analysed by western blotting in the nuclear and cytoplasmic fractions isolated from LO2, HepG2, and Hep3B cell line samples. β-actin and histone H3 were used as the controls for loading and fractionation. g Morphological changes that occurred when LO2, HepG2, HepG2.2.15, and Hep3B cells were treated with TNF-α (40 ng/ml) for 4 h and observed under bright field microscopy. h LO2, HepG2 and Hep3B cells were treated with TNF-α (40 ng/ml) for 4 h, and the expression of ARRB1, p65 and p-p65 was analysed using western blotting. All values are the means ± SD of three separate experiments and were repeated three times. P < 0.01 or P < 0.05 using Student’s t-test