Figure 6. Quantification of telomere G-tail length by hybridization
protection assay in DNA-PKcs knockdown U-2 OS cells. (A) A
schematic of the HPA for telomere G-tail. Non-denatured genomic DNA was
incubated with acridinium ester (AE)-labeled 29-mer telomere HPA probe.
The AE of unhybridized and mis-hybridized probes was hydrolyzed, and
chemilumines-cence from AE of hybridized probes was measured. (B
and C) G-tail length of cells expressing an shRNA control or an
shRNA against WRN was examined in panel B. G-tail length of cells
transfected with siRNA against control (left), siRNA against DNA-PKcs
(middle left), siRNA against DNA-PKcs with pEYFP-WRN (middle right),
or siRNA against DNA-PKcs with pEYFP-WRN (E84A) (right) was examined in
panel C. The G-tail length in the control cells was represented as
100%. Data are represented as mean +/- standard errors of two independent
experiments.
