Figure 3.

The POT1^V29L mutation affects telomeric binding to ssDNA. (a) Western blot of empty (untagged) vector, Myc-POT1 WT and Myc-POT1 mutant with beta-actin serving as an internal control; (b) quantification of telomeric DNA bound to POT1 by ChIP analysis. IgG served as a negative control. Results were normalized to input chromatin. Representative ChIP dot blot is shown with input sample dilutions (left panel). Quantification of ChIP resulted in the bar graph (right panel). Green bar: wild-type; red bar: mutant; (c) results of EMSA showing the decrease in POT1 binding capacity to telomeric ssDNA by the p.V29L substitution; (d) relative telomere length is significantly longer in POT^V29L compared to POT1^WT transfected HEK293T cells after 40 passages. * p < 0.05; ** p < 0.005. The whole blot images can be found in Figure S2 and the POT1 Western blot intensity in Table S3.