Fig. 5: PARP2 STIMULATES BIR-MEDIATED DNA END RESECTION AND STRAND INVASION. | Nature Communications

Fig. 5: PARP2 STIMULATES BIR-MEDIATED DNA END RESECTION AND STRAND INVASION.

From: PARP2 promotes Break Induced Replication-mediated telomere fragility in response to replication stress

Fig. 5

a Model for BIR-mediated fragile telomere formation and their removal after CO-FISH. Representative image of telomere CO-FISH. The Leading strand DNA was hybridized with a red probe and the lagging strand was hybridized with a green probe. White arrows show the fragile telomeres. b, c Quantification of fragile telomeres in PARP1KO and PARP2-FLAG cells after 18 dye and light treatments detected by FISH compared to fragile telomeres detected by CO-FISH on samples derived from BrdU/BrdC-labeled cells, and quantification of leading- and lagging-end telomeres. Each dot represents a metaphase. At least 20 metaphases were analyzed per experiment. Red bars represent mean ± SD from n metaphases analyzed from three independent experiments. P-values were obtained using ordinary one-way ANOVA. d Quantification of the number of pRPA32 foci colocalizing with telomeres per nucleus. Cells were fixed 48 h after the knockdown of BLM with siRNA. Each dot on the graph corresponds to a specific analyzed nucleus. At least 100 cells were counted for each experiment. Red bars represent mean ± SD from n nuclei analyzed from three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA. e Representative images of pRPA32 foci (in red) combined with FISH staining of telomeres (in green) in HeLaFAP, PARP1KO, PARP2KO, PARP1/2KO, PARP2-FLAG, and E558A cell lines. Cells were fixed 48 h after the knockdown of BLM with siRNA. The last row corresponds to zoomed-in squares showing marked pRPA32 foci colocalizing with telomeres. f Quantification of the number of RAD51 foci colocalizing with telomeres per nucleus. Cells were fixed 24 h after the last dye and light treatment (N18). Each dot on the graph corresponds to a specific analyzed nucleus. At least 200 cells were analyzed. Red bars represent mean ± SD. Statistical analysis was performed using ordinary one-way ANOVA. g Quantification of the number of RAD51 foci colocalizing with telomeres per nucleus. Cells were fixed 48 h after the knockdown of BLM with siRNA. Each dot on the graph corresponds to a specific analyzed nucleus. At least 100 cells were counted for each experiment. Red bars represent mean ± SD from n nuclei analyzed from three independent experiments. Statistical analysis was performed using ordinary one-way ANOVA. Source data are provided as a Source Data file.

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