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

(A). Immunohistochemical staining for markers of DNA damage (γH2AX) in TRAMP mice prostate, 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 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 hrs) (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.