Figure 2 | Laboratory Investigation

Figure 2

From: Constitutive Expression and Regulation of Rat Complement Factor H in Primary Cultures of Hepatocytes, Kupffer Cells, and Two Hepatoma Cell Lines

Figure 2

Up-regulation of FH-specific mRNA in HC, FAO, and H4IIE cells after their stimulation with IFN-γ. A, Comparison of IFN-γ-mediated up-regulation in HC, FAO, and H4IIE cells. After incubation of the cells with IFN-γ (100 U/ml), mRNA was extracted and FH-specific mRNA was determined by RT-PCR assays. Only HC respond to IFN-γ with an increase in FH-specific mRNA. The constitutive expression of FH is low in HC, higher in FAO, and absent in H4IIE cells. As a control β actin-specific cDNA was coamplified. B, IFN-γ-dependent up-regulation of the expression of FH-specific mRNA in HC as shown by Northern blot analysis. Left, Gel electrophoresis with staining of extracted mRNA as a control for loading equal amounts of mRNA of both IFN-γ-treated and untreated cells. Right, Northern blot analysis of FH-specific mRNA extracted from HC without stimulation and after stimulation of the cells with IFN-γ (100 U/ml). C, Competitive PCR for the quantification of FH-specific mRNA from IFN-γ-treated and untreated HC. The arrows indicate the points of near equimolarity. The difference between IFN-γ-treated and untreated cells is about 7-fold.

Back to article page