Fig. 2
From: FOXD1 regulates cell division in clear cell renal cell carcinoma
FOXD1 knockout affects expression of cell cycle regulators and alters cellular energetics. a CRISPR/Cas9 knockout targeting strategy for FOXD1. The FOXD1 locus on chromosome 5 shows the single FOXD1 exon with the CDS in dark blue and UTRs in light blue. The placement of the guide RNA (green) relative to the ATG (underlined) is shown below. b Knockout prediction score and percentage of alleles with insertions or deletions (indels) for 82 clones isolated from the CRISPR-edited pool. Chromosome 5 is diploid in 786-O and clones with 50% indel frequency are predicted to be heterozygous, while those with 100% indel frequency are predicted to be homozygous. c Sequencing trace of 786-O versus 786-OFOXD1null. Red arrow indicates nucleotide insertion. Bottom: Predicted protein sequence showing insertion event resulting in modified amino acid sequence (red text) and premature stop (*). d FOXD1 (red) immunofluorescence staining in 786-O versus 786-OFOXD1null, counterstained with DAPI (blue). e Expression of predicted FOXD1 target genes in edited versus parent cells. *p < 0.05