RAP1 deficiency does not affect telomere length in full blown HCC.

(A-B) Mean telomere fluorescence (A) and total nuclear telomere fluorescence (B) in liver sections at 40 weeks post-DEN, 60–65 weeks post-DEN and at the endpoint at 70–100 weeks post-DEN. At 40 weeks the telomere intensity was determined in healthy hepatic tissue. At 60–65 weeks post-DEN before humane endpoint and at the humane endpoint, telomere intensity was determined in HCC. At 60–65 weeks post-DEN, the tumors analyzed presented an area between 10–30 mm2. At the endpoint, the analysis was performed in tumors with an area between 50–500 mm2. (C) Representative Q-fish images showing the telomere FISH (red) in Rap1+/+ and Rap1-/- liver and tumor sections of female mice at different time points. (D-E) Mean telomere fluorescence (D) and total nuclear telomere fluorescence (E) in liver sections at 36 weeks post-DEN and at the humane endpoint of male mice that died of liver tumors at either 55–65 or 65–75 weeks post DEN. At 36 weeks the telomere intensity was determined in healthy hepatic tissue. At the humane endpoint, telomere intensity was determined in HCC. In male mice at humane end points ranging from 55–65 or from 65–75 weeks post-DEN, the tumors analyzed presented an area between 50–350 mm2 and 100–500 mm2, respectively. a.u.f., arbitrary units of fluorescence. Values and error bars represent the mean and SE, respectively. N, number of tumors. Statistical significance was determined by Student’s t test. *p<0.05, **p<0.01, ***p<0.001.