Fig. 6: PARP2 IS REQUIRED FOR POLD3 RECRUITMENT TO TELOMERES. | Nature Communications

Fig. 6: PARP2 IS REQUIRED FOR POLD3 RECRUITMENT TO TELOMERES.

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

Fig. 6

a Representative images of POLD3:TRF2 PLA foci (red) detected in HeLaFAP, PARP1KO, PARP2KO, PARP1/2ko, PARP2-FLAG, and E558A cells after chronic induction of oxidative stress using dye and light. b Quantification of the number of POLD3:TRF2 PLA foci (red) per nucleus detected in HeLaFAP, PARP1KO, PARP2KO, PARP1/2ko, PARP2-FLAG and E558A cells. 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 300 cells were analyzed. Red bars represent mean ± SD. P-values were obtained using ordinary one-way ANOVA. c Representative images of POLD3:TRF2 PLA foci (pink) detected in HeLaFAP, PARP1KO, PARP2KO, PARP1/2ko, PARP2-FLAG, and E558A cells after depletion of BLM with siRNA. d Quantification of the number of POLD3:TRF2 PLA foci (pink) per nucleus detected in HeLaFAP, PARP1KO, PARP2KO, PARP1/2ko, PARP2-FLAG and E558A cells. 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 300 cells were analyzed. Red bars represent mean ± SD. P-values were obtained using ordinary one-way ANOVA. e Immunoblot showing accumulation of PAR from PARP2-FLAG cell extracts treated with PARGi 10 mM for 24 h. Cells were depleted for BLM with 50 nM BLM siRNA for 48 h and were transfected with the POLD3-RFP plasmid. Protein expression was analyzed using anti-BLM and anti-POLD3 antibodies. Actin was used as a loading control. f Representative images of POLD3:TRF2 PLA foci (red) detected in PARP2-FLAG cells after depletion of BLM with siRNA, and inhibition of PARG. g Quantification of the number of POLD3:TRF2 PLA foci (red) per nucleus detected in PARP2-FLAG cells after knockdown of BLM with siRNA and PARG inhibition. 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 two independent experiments. P-values were obtained using ordinary one-way ANOVA. Source data are provided as a Source Data file.

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