Additional file 3 of Alternative splicing of NF-YA promotes prostate cancer aggressiveness and represents a new molecular marker for clinical stratification of patients

Additional file 3: Suppl. Figure S3. CRISPR/Cas9-mediated knock out of endogenous hNF-YA. (A) Schematic representation of CRISPR/Cas9 strategy to knock down human NF-YA. The picture illustrates the sgRNA targeting endogenous hNF-YA (blue line) and the PAM sequence specific for the human gene (red line). Sequence alignment of the reverse complementary strand of exon 2 of human and mouse NF-YA gene is shown. (B) Indel spectrum determined by TIDE analysis on human NF-YA gene in a representative experiment of bulk CRISPR-treated PC3 cells overexpressing murine NF-YAl (left panel) or NF-YAs (right panel). Editing frequencies are shown (p < 0.05). (C) Indel spectrum determined by TIDE analysis on murine NF-YA in CRISPR-treated and GFP-sorted PC3 cells overexpressing murine NF-YAl (left panel) or NF-YAs (right panel). (D) Indel spectrum determined by TIDE analysis on human NF-YA gene in PC3 NF-YAs clone #11 (left panel) or #23 (right panel). Frequencies of editing are reported (p < 0.05) and show biallelic editing.