Fig. 3: Telomere fragility is mediated by PARP2 and its catalytic activity during replication stress. | Nature Communications

Fig. 3: Telomere fragility is mediated by PARP2 and its catalytic activity during replication stress.

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

Fig. 3

a PARP1/2KO cells were transfected with the pCMV-PARP1-3xFlag-WT plasmid, and protein expression was analyzed by immunoblotting using an anti-PARP1 antibody. Actin was used as a loading control. b PARP1/2KO cells were transfected with the pLVX-IRES-puro-PARP2-FLAG plasmid, and its version containing the point mutations, and protein expression was analyzed by immunoblotting using an anti-PARP2 antibody. Actin was used as a loading control. c Modular structures of the ART enzymes PARP2. The amino acid numbers indicate ___domain boundaries. The dark line indicates the point mutation E558A of the catalytic glutamate of the ART ___domain. d Quantification of telomeric signal-free ends detected by FISH in HelaFAP, PARP1/2KO, PARP1-FLAG, PARP2-FLAG, and E558A cell lines after knockdown of BLM with siRNA. Each dot represents a metaphase. Red bars represent mean ± SD from n (35 to 65) metaphases analyzed. P-values were obtained with ordinary one-way ANOVA. e Quantification of fragile telomeres detected by FISH in HelaFAP, PARP1/2KO, PARP1-FLAG, PARP2-FLAG, and E558A cell lines after knockdown of BLM with siRNA. Each dot represents a metaphase. Red bars represent mean ± SD from n (35 to 65) metaphases analyzed. P-values were obtained with ordinary one-way ANOVA. f Quantification of telomeric signal-free ends detected by FISH in HelaFAP, PARP1/2KO, PARP2-FLAG, and E558A cell lines 24 h after the last dye and light exposure (N18). 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 with ordinary one-way ANOVA. P-values were obtained using ordinary one-way ANOVA. g Quantification of fragile telomeres detected by FISH in HelaFAP, PARP1/2KO, PARP2-FLAG, and E558A cell lines 24 h after the last dye and light exposure (N18). 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. Source data are provided as a Source Data file.

Back to article page