Fig. 3: Factors involved in fork reversal are required for nascent strand degradation in PCNA-K164R cells. | Nature Communications

Fig. 3: Factors involved in fork reversal are required for nascent strand degradation in PCNA-K164R cells.

From: Ubiquitinated-PCNA protects replication forks from DNA2-mediated degradation by regulating Okazaki fragment maturation and chromatin assembly

Fig. 3

a RAD51 depletion suppresses HU-induced nascent strand degradation in 293T-K164R cells. The ratio of CldU to IdU tract lengths is presented, with the median values marked on the graph and listed at the top. Asterisks indicate statistical significance (Mann–Whitney test, two-sided). A schematic representation of the fiber combing assay conditions is also presented. A western blot showing RAD51 levels upon siRNA treatment is presented in Supplementary Fig. 5a. b Impact of DNA translocases HLTF, ZRANB3, and SMARCAL1 on HU-induced nascent strand degradation in 293T-K164R cells. The ratio of CldU to IdU tract lengths is presented, with the median values marked on the graph and listed at the top. Asterisks indicate statistical significance (Mann–Whitney test, two-sided). A schematic representation of the DNA fiber combing assay conditions is also presented. Western blots confirming the knockdowns are shown in Supplementary Fig. 5b. c Knockdown of RADX suppresses HU-induced nascent strand degradation in BRCA2-deficient cells, but not in 293T-K164R cells. The ratio of CldU to IdU tract lengths is presented, with the median values marked on the graph and listed at the top. Asterisks indicate statistical significance (Mann–Whitney test, two-sided). A schematic representation of the assay conditions is also presented. Confirmation of RADX knockdown is shown in Supplementary Fig. 5d. Source data are provided as a Source Data file.

Back to article page