Figure EV3. Generation of PTAR1 KO cells by CRISPR‐Cas9 genome editing.

• A
  Generation of PTAR1 KO cell lines by CRISPR‐Cas9 using single‐guide RNA (sgRNA) targeting exon 2 of the human PTAR1 gene. The target sequence and PAM sequence are indicated.
• B
  Sequence data of the genomic region containing the exon 2 of PTAR1. Insertion and deletion mutations are shown in red, and the resulting premature stop codons are underlined. These mutations introduce a stop codon at the amino acid position 42 or 43. HAP1 is a haploid cell line and contains a single allele for PTAR1. nt, nucleotide.
• C
  Immunoblot analysis of WT cells, PTAR1 KO cells, and PTAR1 KO cells stably expressing PTAR1 (KO + PTAR1) using anti‐PTAR1 antibody. Asterisks denote proteins that cross‐react with the antibody.
• D
  Cytosolic localization of Ykt6 in HeLa cells. WT and PTAR1 KO HeLa cells were fractionated into cytosol and membrane fractions, and analyzed by immunoblotting with anti‐syntaxin 5 and anti‐Ykt6 antibody. Syntaxin 5 has two isoforms with different translation initiation sites.
• E
  Prenylation status of Ykt6 in PTAR1 KO HAP1 cells and PTAR1 KO HAP1 cells stably expressing WT PTAR1 or Yk6‐binding defective mutants of PTAR1. The prenylation status of Ykt6 was analyzed by DOC‐PAGE followed by immunoblotting with anti‐Ykt6 antibody. Lower panels show conventional immunoblot analysis of the same samples using the indicated antibodies.

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