Figure 5

Tcstv1/3 knockdown reduces telomere lengths in mouse ESCs and decreases Zscan4 protein levels.

(a) Representative telomere Q-FISH images of Tcstv1/3 KD and control KD ESCs at P8. Telomeres are labeled with telomere PNA probes (green) and chromosomes are labeled with DAPI (blue). Scale bar = 10 μm. (b) Histogram shows distribution of relative telomere length expressed as TFU (telomere fluorescence unit) by telomere Q-FISH analysis. Average telomere length is shown as mean TFU ± SD. P value, compared to control KD ESCs by t-test. (c) Telomere restriction fragment (TRF) analysis showed decreased telomere lengths in Tcstv1/3 KD ESCs compared with control KD ESCs at P8. (d) Confirmation of Zscan4 expression levels in Tcstv1/3 KD and control ESCs by western blot. Zscan4 band is indicated by black arrow. (e) Relative Zscan4 protein quantity normalized to β-actin by ImageJ software. n = 4; *P < 0.05, **P < 0.01, compared to controls.