Figure 2
Effect of L75P-ApoA-I on ANG expression and cellular localization. (a) qRT-PCR analysis of ANG mRNA. Cells were cultured in the presence (black bars) or absence (grey bars) of fetal bovine serum (FBS) for 48 h. Total RNA was extracted and analyzed with probes specific for ANG and ACTB mRNAs. ΔΔCt method was used. The amount of ANG mRNA was normalized to the endogenous actin and each ΔCt was then normalized to the value of vector control transfectants. Data shown are the means±S.D. of three independent experiments. (b) Immunoblot analysis of cytoplasmic and nuclear ANG. Cells were cultured in the presence or absence of FBS for 48 h. Cytoplasmic and nucleolar fractions were obtained by subcellular fractionation. A total of 100 μg proteins was loaded in each lane. (c) Ratio of cytoplasmic versus nucleolar ANG. (d) Relative abundance of cytoplasmic ANG normalized to β-actin. (e) Relative abundance of nucleolar ANG normalized to nucleophosmin B23. Data shown in c–e are the means±S.D. of three independent experiments
