Fig. 6: cGAS inhibits alt-NHEJ by suppressing Polθ transcription in a Ku80-dependent manner.
From: The cGAS-Ku80 complex regulates the balance between two end joining subpathways
A RT‒qPCR analysis of the relative Polθ mRNA level in Ku80-overexpressing HCA2-hTERT cells. B RT‒qPCR analysis of the relative Polθ mRNA level in HCA2-hTERT cells with Ku80 depletion. C Analysis of Polθ promoter activity in cells overexpressing Ku80 using the PPolθ-Luc reporter. D Analysis of alt-NHEJ efficiency in Ku80-overexpressing HCA2-hTERT cells using EJ2-GFP reporters. E Analysis of alt-NHEJ efficiency in HCA2-hTERT cells with Ku80 depletion using EJ2-GFP reporters. F ChIP assay showing the recruitment of Ku80 to Polθ promoter regions. Vectors expressing Ku80-HA were transfected into HEK293T cells. Twenty-four hours post-transfection, cells were harvested for a ChIP assay with an antibody against HA. G RT‒qPCR analysis of the relative Polθ mRNA level in HCA2-hTERT cells overexpressing cGAS and/or Ku80. H ChIP assay showing that depleting Ku80 by siKu80 impaired the enrichment of cGAS at the Polθ promoter in HEK293T cells. I RT‒qPCR analysis of the relative Polθ mRNA level in control or cGAS-overexpressing HCA2-hTERT cells with or without Ku80 depletion. Data are representative of 3 independent experiments. Data are expressed as the mean ± S.D. values. Student’s t test or one-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s., not significant.
