Figure 5
From: Immortal Activated Human Hepatic Stellate Cells Generated by Ectopic Telomerase Expression
Gene expression patterns in activated human HSCs and telomerase-positive HSCs. A, Microarray analysis of gene expression in activated human HSCs and telomerase-positive HSCs was performed. The values of all genes in activated human HSCs (x axis) and telomerase-positive HSCs (y axis) were compared using scatter plot analysis. The upper and lower diagonals represent 2-fold higher gene expression in telomerase-positive and activated human HSCs, respectively. B, Uninfected and vector control–infected HSCs, telomerase-positive polyclonal HSCs, and HSCs from clone 14 were cultured on plastic. Total RNA (5 μg) was subjected to RNase protection assay with riboprobes specific for human collagen α1(I) and human GAPDH mRNA. Migration of the protected bands is indicated. C, Western blot analysis was performed using whole cell extracts obtained from uninfected HSCs, vector control–infected HSCs, telomerase-positive polyclonal HSCs, and HSCs from clone 14. Proteins (10 μg) were separated in a 12% SDS-PAGE, transferred to a nitrocellulose membrane, and immunoblotted for αSMA using monoclonal anti-smooth muscle α-actin antibody.
