Fig. 1

Generation of KRT14-T2A-GFP leader cell lines. A Schematic illustration of the KRT14-T2A-GFP construct. B COV362.4, SKOV3, and OVCAR4-KRT14-T2A-GFP cells, LCs marked by GFP, all cells marked by RFP. C KRT14-T2A-GFP lines were fixed and stained with a polyclonal KRT14 antibody and showed positive co-staining. Scale bar represents 300 μm. D Homozygous KRT14-T2A-GFP cells were transfected with constructs: “KRT14^KO” - targeting exon 1 of the KRT14 gene or the “non-targeting” lentiCRISPR V1 control alone. Cells were imaged at 4X magnification for GFP and RFP 72-h post transfection. E LC^GFP+ counts in KRT14^KO CRISPR targeted lines demonstrated a significant reduction of GFP expression compared to the non-targeting lentiCRISPR V1 control by unpaired t-test and Welch’s correction (p < 0.001) (n = 4 replicates from one representative experiment). Scale represents 100 μm