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. 2021 Jun 15;40(14):e106355. doi: 10.15252/embj.2020106355

Figure EV1. (related to Fig 1). Allele sequences in CRISPR/Cas9‐derived PRIMPOL KO U2OS cells, cell synchronization, and markers of DNA damage induced by MMC.

Figure EV1

  1. Flow cytometry profiles of DNA content (PI) in the synchronized cell cultures used for PrimPol IP and mass spectrometry. The percentage of cells in each phase of the cell cycle is indicated.
  2. Genomic sequences derived from WT and KO alleles in U2OS‐PRIMPOL KO cells obtained with CRISPR/Cas9. The Cas9 guide RNA sequence is marked in red. As the 4q35.1 chromosome locus is amplified in U2OS cells, three PRIMPOL alleles were targeted to achieve a full KO.
  3. Immunoblots show the levels of the indicated proteins in whole cell extracts from the same samples used in Fig 1C. The position of the ubiquitylated form of FANCD2 (Ub‐FANCD2) is indicated. Tubulin is shown as a loading control. Protein names in black and blue indicate the two different gels used.

Source data are available online for this figure.