(A,B) Parental HepG2 or cells expressing NTCP-WT or NTCP-N5,11Q were treated with 0.1% DMSO, Bafilomycin (10nM/24h), MG132 (20μM/6h) or a combination of Bafilomycin (10nM/24h) and MG132 (20μM/ for the last 6h). (A) Immunofluorescence microscopy of NTCP-WT and NTCP-N5,11Q. in which NTCP was visualized using anti-HA antibody (green) and Hoechst (blue) was used to visualize the nucleus. (B) Results of TCA uptake in parental HepG2 cells (grey) and HepG2 cells expressing NTCP-N5,11Q (black). (C) Results of TCA uptake in HepG2 cells expressing NTCP-WT (grey) or NTCP-N5,11Q (black). (B,C)The uptake capacity of the NTCP was determined as pmol TCA uptake/well. Bars represent the mean +/- sd of three independent experiments, each performed in quadruplicate. *Indicates significant different from DMSO treated (p <0.05). # indicates significant different between the cell-lines in the same treatment (p<0.05).