Extended Data Fig. 5. Creation and validation of NC8 cells knocked out for LDLR.
a, Gating strategy and PE intensity from α-LDLR-PE staining are shown. α-LDLR-PE staining was evaluated on single cells. b, Sorting results from bulk NC8 iPSC after Cas9 LDLR editing. PE intensity from α-LDLR-PE staining is shown for cells targeted with an LDLR guide RNA or for unmodified control cells. Event densities were smoothened and are displayed as absolute counts or as counts normalization to the mode. c, Qualitative flow-cytometry result of selected clones stained with an α-LDLR-PE antibody. Shown is the PE intensity from α-LDLR-PE staining from gated single cells of LDLR- or LDLR+ iPSC clones. d, Flow-cytometry result of the studied LDLR-KO or wild-type LDLR iPSC clones (clone 4, clone 10). Shown is the overlayed mode-normalized density of PE intensity from α-LDLR-PE staining for clone 10 and 4. The legend percentages indicate the fraction of α-LDLR-PE negative stained single cells. e, Sanger sequencing of PCR product from the LDLR genomic locus CRISPR-Cas9 editing site for NC8 iPSC clones 10 and clone 4 are shown. The sgRNA spacer, PAM and expected Cas9 editing site (3 base-pairs downstream of PAM sequence) are shown above the sequencing traces.