Figure 4
From: Expression of HSP47, a collagen-specific chaperone, in normal and diseased human liver
(a) RT-PCR detection of HSP47 mRNA and corresponding levels of immunoreactive HSP47 in human livers. Total RNA was isolated from human livers and reversed transcribed. The cDNA products were amplified (30 cycles) using specific primers for HSP47 and GAPDH as described in the Materials and methods. Western analysis was performed as described for Figure 2 and in the Materials and methods. RNA and protein samples were obtained from the same livers for these measurements. The first lane is a ladder (top and middle panels) or molecular weight markers (bottom panel). Control, samples from normal donor livers; ALD, cirrhosis due to alcoholic liver disease; HCV, cirrhosis due to chronic hepatitis C infection. (b) Real-time quantitation of HSP47 mRNA in control and cirrhotic human livers. Total RNA from the same livers used for the semiquantitative RT-PCR shown above was reverse transcribed and subjected to real-time PCR using the TaqMan system as described in the Material and methods. Abundance of HSP47 mRNA was determined using the comparative cycle threshold method. Data are means±s.e.m.; the difference between control and cirrhotic livers was not significant (P=0.08).
