Research Paper Volume 14, Issue 7 pp 3259—3275

Figure 2. Cholesterol increased LAP3 expression, which is mainly localized in the nucleus of LO2 cells. (A–C) After LO2 cell line treatment with 150 μM CHO, 200 μM PA, 5 μM LPS or 10 ng/ml TNF-α for 24 h, the mRNA expression of lap3 was detected by RT-qPCR (A), the protein expression of LAP3 was detected by western blotting (B) and normalized by β-actin (C). (D and E) LO2 cell line treated using different concentration 0, 50,100, 150 and 250 μM of CHO for 24 h respectively and the protein expression of LAP3 was detected by western blotting (D) and normalized by β-actin (E). (F–J) LO2 cell line treated with 150 μM of CHO for the indicated time points, the protein expression of LAP3 was detected by western blotting (F) and normalized by β-actin (G), and further validated by IHC (H) and IF (J), respectively. (I and K) Statistics for IHC and IF by Image pro plus. Data are expressed as means ± SD from three independent experiments. ^*p < 0.05; ^**p < 0.01; ^***p < 0.001. An unpaired t-test and one-way ANOVA were applied to determine the statistical significance. Abbreviations: CHO: cholesterol; PA: palmitic acid; IHC: immunohistochemistry; IF: immunofluorescence.