Fig. 3: Nuclear actin polymerization is required for active fork slowing upon mild genotoxic stress. | Nature Communications

Fig. 3: Nuclear actin polymerization is required for active fork slowing upon mild genotoxic stress.

From: Nuclear actin polymerization rapidly mediates replication fork remodeling upon stress by limiting PrimPol activity

Fig. 3: Nuclear actin polymerization is required for active fork slowing upon mild genotoxic stress.The alternative text for this image may have been generated using AI.

a–d DNA fiber analysis of U2OS cells. a Top: Schematic CldU/IdU pulse-labeling protocol used to evaluate fork progression upon 100 nM CPT or 20 nM ETP. 100 nM LatB, Swi, CK666 or CK869 were added 10 min prior to CPT or ETP and retained during the IdU labelling. Bottom: Representative DNA fiber images. Scale bar = 5 μm. b–d, f IdU/CIdU ratio is plotted for a minimum of 100 forks (indicated as black dots) from a single representative experiment. Red line indicates the median. See Supplementary Fig. 3b–d, h for compiled repetitions (n = 3). Statistical analysis: two-tailed Mann–Whitney test. e, f DNA fiber analysis of RPE-1 cells stably expressing doxycycline inducible BFP-NLS or NLS-BFP-ActinR62D. e Top: schematic of the CldU/IdU pulse-labeling protocol used to evaluate fork progression upon 100 nM CPT. Doxyclicline (Dox) was added 24 h before CldU/IdU pulse-labeling. Bottom: representative DNA fibers images. Scale bar = 5 μm. Source data are provided as a Source Data file.

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