Functional characterization of CRISPR-Cas9-edited monoclonal melanocyte cell lines
(A) Genomic DNA sequences of (multiple) monoclonal cells homozygous for rs117132860-G (WT), heterozygous monoclonal cells with one wild-type allele and second copy with a 16 bp deletion including the SNP (HT), and two monoclonal clones that each have both alleles harboring a short deletion encompassing the AHR-binding motif (KO1 and KO2).
(B) Crystal violet staining of rs117132860-WT1 and rs117132860-KO2 cells not treated with UVB (24 h), 72 h after UVB treatment, and at day 7 after UVB exposure, followed by a zoomed-in image of day 7.
(C) FACS analysis of the same WT and KO cells at day 7 following UVB treatment. Forward-scatter and side-scatter image indicates a mixed population, and BrdU and 7-AAD staining characterize the cell cycle status of each population. The images shown here are from a representative experiment from three biological replicates. Additional WT and KO clones were assessed similarly.