Figure 2.

(A) Induction of Zf9 during stellate cell activation in vivo; Western blot analysis of freshly isolated stellate cell lysates from a normal rat and a rat treated 3 hr earlier with CCl₄. A single protein of ≈46 kDa is identified. The blot was reprobed with anti-phosphatidylinositol 3-kinase. The induction after CCl₄ represents a 3-fold increase. A similar induction was reproducible in three experiments. (B) Increased biosynthesis of Zf9 during stellate cell activation in vivo. Pooled stellate cells from either two normal rats or two rats administered CCl₄ were plated in methionine-free medium containing [³⁵S]methionine and harvested at 45, 90, and 190 minutes or after 190 minutes with [³⁵S]methionine followed by a 20-hr chase in unlabeled medium. [³⁵S]Zf9 was purified by immunoaffinity chromatography by using a column containing anti-Zf9. SDS/PAGE autoradiography of eluted protein confirmed specific incorporation of [³⁵S]methionine into Zf9 (not shown). Data is depicted here as specific [³⁵S]methionine incorporation into protein from normal and activated stellate cells, normalized for cell number.