Fig. 2

The SETDB1-binding region, but not the FNIII domain, within ATF7IP is required for normal localization of SETDB1. a IF analysis shows that exogenous expression of 3xFLAG-ATF7IP WT and dFNIII mutant, but not dSETDB1 mutant, rescues the abnormal localization of SETDB1 in Atf7ip KO mESCs. Representative images are shown, and the quantitative analyses are shown in F–H. Scale bar: 10 µm. b SETDB1 (left) and 3xFLAG-ATF7IP (right) signals in the nucleus that was determined by DAPI staining were calculated. The mean from three independent experiments is shown as a bar graph with jittered points indicating the average % intensity of each experiment. Over 100 cells were analyzed per sample per experiment. NS (not significant): P > 0.05, ***P < 0.001 versus “ + Empty” group by Dunnett’s test. c, d SETDB1 (left) and 3xFLAG-ATF7IP (right) foci in the nucleus that was determined by DAPI staining were calculated. Violin plot for their foci numbers from all the cells analyzed are shown (c). The mean from four independent experiments is shown as a bar with jittered points indicating the average number of each experiment (d)