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. Author manuscript; available in PMC: 2010 Jun 21.
Published in final edited form as: Lab Invest. 2008 Dec 29;89(2):209–221. doi: 10.1038/labinvest.2008.127

Figure 1.

Figure 1

The effect of PTK/ZK on HSC activation in culture. (a) Expression of HSC activation marker α-SMA was significantly inhibited by PTK/ZK. Isolated HSCs were incubated with PTK/ZK for 24 h at the concentration of 5 or 10 μM, and labeled with α-SMA antibody for flow cytometry analysis. The percentage of α-SMA-positive cells was significantly decreased followed by PTK/ZK treatment. Data are mean±s.e. of three different experiments. (b) Effect of PTK/ZK on HSC proliferation. HSCs were cultured for 72 h in the presence of various concentrations of PTK/ZK. Proliferation was measured by BrdU incorporation assay. Results were representative of three experiments and each concentration was repeated six times in each experiment. (c) PTK/ZK inhibited migration of HSC measured by using Invasion Chamber as described in Materials and Methods section. HSCs were incubated in serum free medium for 24 h in the upper compartment of the chamber and were tested for migration in the lower compartment in the present of various concentrations of PTK/ZK. Cells migrated through to the lower compartment were quantified by cell counting. Values are expressed as a percentage of untreated control from three independent experiments. (d) PTK/ZK inhibited cytoskeleton rearrangement in activated HSCs. Fluorescence microscope shows changes in the cytoskeletal organization of rat HSCs after 24 h culture in the presence of PTK/ZK (10 μM). Fluorescent conjugates of phalloidin were used to label actin filaments. (e) HSCs were treated with incremental concentrations of PTK/ZK for 24 h. Analyzed by western blot, expression of collagen I was significantly inhibited by PTK/ZK. (f) Quantification of mRNA levels of TIMP-1. Total RNA was isolated from HSCs using RNeasy Mini kit, and TIMP-1 gene expression was measured by real-time PCR. PTK/ZK significantly suppressed TIMP-1 expression. TIMP-1 values were normalized to 18S mRNA values. Data are expressed as mean±s.e., *P<0.05.