Figure 3.
Sustained E2F Transcription Is Necessary for Checkpoint Functions
(A) DNA fiber analysis schematic and representative images of individual fibers, HEK293 T-Rex E2F6 cells. Bar graphs of green track length, 2 hr HU −/+ E2F6 (Doxy 2 μg/ml). p, significantly longer tracks with E2F6; Student’s t test.
(B) Schematic and representative images, HEK293 T-Rex E2F6 cells. Bar graphs of green track length, 4 hr chase in HU −/+ E2F6 (Doxy 2 μg/ml). p, significantly shorter tracks with E2F6; Student’s t test.
(C) Bar graph of % cells with RPA2 foci at times shown after release from 16 hr HU −/+ E2F6 (Doxy 2 μg/ml). n = 3; >130 for each; error bars = SEM. ∗∗∗p < 0.001; ∗p < 0.05 with ANOVA and Tukey’s.
(D) Representative images for (C). The scale bar represents 10 μm and is the same for all.
(E) Western blot of chromatin preparation, RPE1 TetON E2F6 cells, 7 hr, treatments shown. E2F6, Doxy 2 μg/ml.
(F) Western blot of WCE, RPE1 TetON E2F6 cells, 7 hr, treatments shown. E2F6, Doxy 2 μg/ml.