Met and HGF expression in primary and immortalized HSC.
Total RNA was extracted from primary rat HSC after the third, fifth and seventh day of cell culture (3d, 5d, 7d) and from myofibroblastic HSC-T6 cells. Subsequently, c-met and HGF expression was determined (A–C). The c-met and HGF transcript levels were quantified by Real-Time PCR and normalized using HPRT as house-keeping gene (A–C). The expression of c-met increased during the differentiation process of primary HSC and achieved the highest level in the immortalized cell line HSC-T6 representing a myofibroblastic phenotype , whereas HGF levels were shown to be opposite (A, B). Primary HSC at the 3rd culture day (prim. HSC) and immortalized HSC (HSC-T6) were either untreated (−) or stimulated with TGF-β (+) and RNA levels of c-met and HGF were analyzed by Real-Time PCR (C). The Met protein expression in HGF and TGF-β stimulated HSC-T6 (+HGF, +TGF-β) in comparison to untreated HSC-T6 cells (control) was shown by Western blot analysis (D).