Figure 2.
DACH1 inhibition of cellular proliferation requires c-jun. (A) Western blot analysis of c-junfl/fl 3T3 cells transduced with adenovirus expressing Cre recombinase. Cells were transduced with the MSCV–DACH1 vector as indicated. Western blot analysis shows Ad-Cre excision of c-jun abrogated c-jun expression. Antibodies are to cyclin D1, cyclin A, and β-tubulin. Cellular proliferation assessed by MTT assay (B) and by cell counting of DACH1-transduced cells (C) shown in A. (D) Proliferation assay of 3T3 Tet-c-Jun cells, in which 2 μg/ml doxycycline for 48 h was used to induce c-Jun protein level. (E) NIH3T3 cells transduced with pLRT-GFP, pLRT-DACH1, or pLRT-DACH1 and pLRT-c-Jun expressing vector. Cellular number was determined daily for 6 d with 2 μg/ml doxycycline. c-Jun expression reversed DACH1-mediated inhibition of cellular proliferation (p < 0.01).