Where indicated, data were subjected to a two-tailed Student’s t test to resolve statistical significance. The measured TFI values of GFP molecules and single-labeled mRNA transcripts were plotted into histograms and fitted with a Gaussian function according to the formula Embedded Image to determine the mean value xc of the intensity distribution in OriginPro 8. The mean TFI value for single GFP molecules was related to the mean single transcript intensities via conversion of the exposure time at identical imaging settings. The comparison of DNA ligase IV knockdown and control live cell experiment datasets was performed using Fisher’s exact test for 2 × 2 contingency tables and a χ2 test for contingency tables, both at a level of significance of P = 0.05.

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