Fig. 5

Inhibition of deneddylation function of the CSN complex resulted in increased levels of SMC6 localization to laser induced DNA damage. (a) Western blot assessing proteins within the soluble and chromatin bound fractions extracted from cells treated with vehicle control (DMSO), MLN4924 (neddylation inhibitor) and CSN5i-3 (deneddylation inhibitor). A portion of cells from each condition were exposed to UV radiation (75 J/m²) for 30 min to enhance chromatin recruitment of the CUL4 complex. (b) Representative images of cells exposed to laser-induced DNA damage following treatment with vehicle control or CSN5i-3. (c) Box and whiskers plot showing the quantification of SMC6 signal intensity at the site of laser-induced DNA damage compared to average signal intensity within the undamaged regions of the nucleus following treatment with vehicle control or CSN5i-3. P-value of < 0.0001 was determined from a two-tailed Mann Whitney t-test. Whiskers represent the minimum and maximum measurements. Ends of the box are the upper and lower quartiles, and the median is marked by a vertical line inside the box. Size bars = 10 μm