Figure 2
AMG 900 inhibits p-Histone H3 in COLO 205 tumors and mouse skin (hair follicle, epidermis) in a dose-dependent manner measured by Laser Scanning Cytometry (LSC). Mice bearing established tumors were orally administered a single dose of vehicle alone or AMG 900 (as described in Figure 1). Mouse skin specimens (a right- and left-flank punch biopsy from each animal) were collected three hours after treatment (n = 10 per treatment group) and processed for p-Histone H3 analysis by LSC. Small intestine was also collected as a positive staining control for p-Histone H3. Images were captured on a LSC (iCys, CompuCyte) using a 20× objective. See Materials and Methods Section for tissue and tumor section immunofluorescence staining procedure and Additional file 2 Figure S2A for the random sampling contour strategy. (A) Representative scan fields of mouse tissues (skin, hair follicle, small intestine) and COLO 205 tumor showing p-Histone H3 positive objects (red or pink). Nuclei were counterstained with Hoechst 33342 (blue). An average p-Histone H3 object count (per mm2) was determined from three scan fields for each tissue section. Column graphs represent p-Histone H3 positive object for skin (B) and COLO 205 tumor (C) (mean + SE). Statistically significant inhibition of p-Histone H3 compared with vehicle-treated control (*P <0.0001; **P =0.0005).