Figure 5. Microarray analysis of HCECs for LAT1 transporter protein, CD74, HLA-DR, and HLA-DRB1 and western blot analysis. A:All results shown are normalized against the control (unstimulated HCECs). We could demonstrate a fourfold upregulation of the LAT-1 transporter system upon stimulation with IFN-γ (together with a 2.7 fold upregulation of the class II surface antigens) and a 3.85 fold upregulation following stimulation with the cocktail of IFN-γ and TNF-α. However, the stimulation with TNF-α alone only caused a 1.9 fold upregulation. B: Semi-quantitative western blot analysis of LAT1 protein: 1=protein ladder (250 D-10 kDa (Bio-Rad), 2=unstimulated HCECs, 3=HCECs stimulated with IFN-γ, 4=HCECs stimulated with IFN-γ+TNF-α, 5=HCECs stimulated with IFN-γ, TNF-α+Prostaglandin E2 (as a control to antagonize the stimulatory effects of IFN-γ+TNF-α), 6=HCECs stimulated with IFN-γ, TNF-α+Prostaglandin E2+1-MT, 7=HCECs stimulated with TNF-α, 7=HUH-7 liver cell line as control. C: The bottom panel shows the exemplary clustering analysis of our Affymetrix assay. D: Upregulation of LAT1 following stimulation with IFN-γ, TNF-α and IFN-γ, TNF-α was detected by quantitative real-time PCR. The data shown are normalized against the copy numbers determined from unstimulated HCECs.