Species specific fitc conjugated secondary antibodies

Cells were trypsinized and seeded on coverslips (Thermo Fisher Scientific, Lafayette, CO) in 24-well plate overnight. The next day cells were irradiated with single dose and allowed to recover for indicated time prior to analysis. Immunostaining was performed according to a previous report [15 (link)]. Anti-γH2AX (#9718, 1:200; Cell signaling Technology), anti-Rad51 (ab63801, Abcam) and species-specific FITC-conjugated secondary antibodies (1:500; Invitrogen) were used. Gray level images were acquired under a laser scanning microscope (Axio Imager.Z2, Carl Zeiss Co. Ltd.). Images were captured using the AxioVision Rel.4.6 computerized image analysis system. At least 50 cells were counted for quantification of γ-H2AX foci.

Cells were seeded on coverslips (Thermo Fisher Scientific, Lafayette, CO) and cultured in 24-well plate overnight. Immunostaining was performed according to a previous report [16 (link)]. Anti-β-catenin (1:200, Cat#9562, Cell signaling technology, Danvers, MA, USA) antibody and species-specific FITC-conjugated secondary antibodies (1:500; Invitrogen, Carlsbad, CA, USA) were used. Image-Pro Plus 6.0 (Olympus) was used for image analysis.