Figure 2. Expression of DNA damage-induced mediator proteins at 4 hr and 24 hr in HT29 cells following inhibition of GLI1/GLI2 by GANT61 (20 μM).

A: Localization of γH2AX, MDC1 and NBS1 nuclear foci in single cells and their co-localization was determined after GANT61 administration. After cells were fixed, permeabilized, treated with the respective antibodies, and stained, four-color image acquisition was performed by confocal microscopy and post-processing analysis of the images was as described in Materials and Methods. B: Western analysis of the expression of total and activated forms (γH2AX, p-ATM, p-MDC1, p-NBS1^Ser343) of mediator proteins was examined after treatment of HT29 cells with GANT61.