Fig. 1: Creating and characterizing a cell line expressing mAID-tagged polE1.
From: The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells
a Wild-type U2OS, homozygous mAID-KI clones 16 and 1.6 were treated for 24 h with doxycycline, auxin, or both, as indicated, western blots of total cell lysates are shown. b Equal numbers or wild-type U2OS, or homozygous mAID-KI clones 16 or 1.6, were seeded on 60 mm dishes and treated with DMSO or dox/aux for 72 h. The data are depicted as mean + SD from n = 3 independent experiments. c Clone 16 was treated with doxycycline overnight (16 h), auxin was added for the indicated times; western blots of total cell lysates are shown. d–f Wild-type U2OS, homozygous mAID-KI clone 16 were treated for 24 h with DMSO or dox/aux, 10 µM EdU was added for the last 30 min of treatment. d Flow cytometry plots showing EdU incorporation and DNA content (7-AAD staining) are shown. e, f Quantification of the flow cytometry data is shown—mean + SD from n = 3 independent experiments. Paired t-test was used for statistical analyses, p values are shown where they are statistically significant. e EdU+ cells were quantified based on EdU signal being above G1 and G2 levels. f Cells were assigned to G1 based on DNA content of 2n and being EdU negative. Source data are provided as a Source data file.
