Fig. 3

A Standard curve and linear relationship for q-TRAP. The Ct values (± s.d.) of the standard control (MCF-7) were plotted against log [protein] to calculate the linear equation. The Y-intercept and the slope values from the equation are used to quantify the RTA of samples. B Telomerase activity as measured by q-TRAP assay. For q-TRAP quantification, the RTA for a sample was calculated based on the equation obtained from the standard curve y = -2.529 x + 21.38; RTA = 10^ [(Ct sample x Yint)/slope]. Sample results are plotted as percentage to control. C Representative WB analysis of TERT in NCI-H929 cell lines 48 h after hit 17 treatments. D Representative WB analysis of histone H3, H4 and try-methylation of H3 at lysine 9 (H3K9me3), H3 at lysine 27 (H3K27me3) and H4 at lysine 20 (H4K20me3) in NCI-H929 cell lines 48 h after hit 17 treatments. E Representative images showing detection of heterochromatin trimethylation markers by immuno-FISH in hit 17 and control (DMSO) in NCI-H929 cell lines. Interphase cells were stained with anti-H3K9me3 (green), anti-H3K27me3 (green), anti-H4k20me3 (green) and telomeric Cy3-labeled probe (red). Nuclei were counterstained with DAPI (blue). Scale bar, 10 μm. ** P <0.01; *** P <0.001