Extended Data Figure 8: Induction of Pd-l1 3′-UTR deletions and inversions in mouse cell lines using the CRISPR-Cas9 system.
From: Aberrant PD-L1 expression through 3′-UTR disruption in multiple cancers
a, Positions of targeting sgRNAs used for CRISPR-Cas9-mediated disruption of Pd-l1 3′-UTR are indicated by arrows. b, Pd-l1 surface expression in EG7-OVA cells transfected with Cas9 and no, single, or pairwise sgRNAs. Representative of three independent experiments. c, d, PCR detection of the Pd-l1 3′-UTR deletion (c) or inversion (d) breakpoint junction from EG7-OVA, P815, and B16-F10 cells in which Cas9 was expressed without (parental) or with no sgRNA (mock), or a pair of Pd-l1 sgRNAs. e, Sequence chromatogram of the detected Pd-l1 3′-UTR deletions from sgPd-l1-transfected EG7-OVA, P815, and B16-F10 cells. f, g, Pd-l1 exon 4 mRNA expression (RPKM) was calculated from the RNA-seq data for EG7-OVA, P815, and B16-F10 cells in which Cas9 was expressed without (parental) or with no sgRNA (mock), or a pair of Pd-l1 sgRNAs (f). RNA-seq reads within the Pd-l1 gene were visualized by IGV (g).
