Extended Data Figure 9: Induction of PD-L1 3′-UTR deletions and inversions in human cell lines using the CRISPR-Cas9 system.
From: Aberrant PD-L1 expression through 3′-UTR disruption in multiple cancers
a, PCR detection of the PD-L1 3′-UTR deletion breakpoint junction from T2 cells in which Cas9 was expressed without (parental) or with no sgRNA (mock), or a pair of PD-L1 sgRNAs. b, Sequence chromatogram of the detected PD-L1 3′-UTR deletions from sgPD-L1-transfected HEK293T and T2 cells. c, PCR detection of the PD-L1 3′-UTR inversion breakpoint junction from HEK293T, T2, and PC-9 cells in which Cas9 was expressed without (parental) or with no sgRNA (mock), or a pair of PD-L1 sgRNAs. d, Sequence chromatogram of the detected PD-L1 3′-UTR inversions from sgPD-L1-transfected HEK293T, T2, and PC-9 cells. e, Visualization of RNA-seq reads within the PD-L1 gene for T2 and PC-9 cells in which Cas9 was expressed without (parental) or with no sgRNA (mock), or a pair of PD-L1 sgRNAs. f, Flow cytometric analysis of PD-L1 surface expression in parental or sgPD-L1-transfected PC-9 cells stimulated with IFN-γ (100 or 300 U ml−1) for 48 h. Representative of three independent experiments.
