Figure 4.

Parental Histone Redistribution Is Controlled by the Repair Factor DDB2

(A) Scheme representing the main repair factors involved in UVC damage detection in the global genome NER pathway. Microscopy images show the distribution of parental H3.3 (red) 15 min after UVC laser damage in U2OS cells stably expressing H3.3-SNAP and GFP-XPC treated with the indicated siRNAs (siLUC: control). siRNA efficiencies were verified by western blot. The red fluorescence loss measured in damaged areas is normalized to before laser (n cells scored in two independent experiments).

(B) Distribution of parental H3.3 (red) 15 min after UVC laser damage in U2OS cells stably expressing H3.3-SNAP and GFP-XPC or GFP-DDB2. Expression levels of exogenous XPC and DDB2 relative to the endogenous proteins are shown on the western blot. Red fluorescence loss is measured in irradiated areas relative to before laser and the area of fluorescence loss is marked by GFP-tagged NER factors.

(C) Distribution of parental H3.3 (green) upon tethering of mCherry-LacR (LacR) or mCherry-LacR-DDB2 (LacR-DDB2) to the LacO array in U2OS LacO cells stably expressing H3.3-SNAP. The area of the LacO array and green fluorescence at the LacO array are displayed on the boxplots (n cells scored in two independent experiments). White arrowheads, irradiated areas or LacO array. Scale bars, 10 μm.

See also Figures S4 and S5.