Fig. 5. Prostatic Dach1 governs the DNA damage response in TRAMP mice.

A Immunohistochemical staining for markers of DNA damage (γH2AX) in TRAMP mice prostates, with (B) Quantitation data shown as mean ± SEM for percentage of γH2AX positive cells (P = 2.07 × 10⁻⁷ by Student’s t test) (n = 19 for Dach1^wt/wt mice, 3 separate mice, 6 views for two mice, 7 views for one mouse) (n = 23 for Dach1^fl/fl mice, 4 separate mice, 7 views for two mice, 6 views for one mouse, 3 views for one mouse) (left panel). Data are shown as mean ± SEM for relative intensity of γH2AX (n = 16 for Dach1^wt/wt mice, 3 separate mice, 5 views for two mice, 6 views for one mouse) (n = 20 for Dach1^fl/fl mice, 4 separate mice, 6 views for two mice, 5 views for one mouse, 3 views for one mouse) (P = 2.2 × 10⁻⁵ by Student’s t test) (right panel). C Western blot of Dach1^+/+ or Dach1^−/− 3T3 cells. D γH2AX immunofluorescent staining of Dach1^+/+ or Dach1^−/− 3T3 cells, with (E) quantitation shown as mean ± SEM, (n = 20 separate cells). F LNCaP cells transduced with shDACH1, treated with arsenic trioxide (ATO 1 h) (10 μM) and (G) quantitation shown as mean ± SEM (n = 20 cells). H LNCaP cells stably transduced with control vector or shDACH1 were treated with ATO (1 μM) for the time points indicated. Western blotting was conducted for γH2AX, with quantitation of a representative experiment shown in (I). J LNCaP cell line stably expressing doxycycline-inducible DACH1 were analyzed for the abundance of γH2AX and other proteins as indicated. GAPDH was used as a protein loading control. S.E. short exposure, L.E. long exposure.