Normal surface HVEM expression in mHVEM mutant mice. (A) Schematic of nucleotide sequences of the HVEM gene in mHVEM mutant mouse strains (G72R/V74A: mHVEM−BT/160, loss of BTLA and CD160 binding; H86D/L90A: mHVEM−LIGHT, loss of LIGHT binding) that were generated by CRISPR-Cas9 editing of exon 3 of the Tnfrsf14 locus. Red letters indicate mutated nucleotides. Green letters indicate protospacer adjacent motif (PAM) sequence. Blue letters indicate mutated amino acids. Black box shows restriction enzyme sites. (B) HVEM surface expression level of splenic CD4+ T cells, ILCs (CD3−Lin−CD90.2+), and iNKT cells (TCRβ+, CD1d tetramer+) from the indicated mice were determined by flow cytometry. HVEM-knockout mouse is a negative control for HVEM staining. KI, KI allele. ssDNA, single-stranded DNA.
Figure S3.

Normal surface HVEM expression in mHVEM mutant mice. (A) Schematic of nucleotide sequences of the HVEM gene in mHVEM mutant mouse strains (G72R/V74A: mHVEM−BT/160, loss of BTLA and CD160 binding; H86D/L90A: mHVEM−LIGHT, loss of LIGHT binding) that were generated by CRISPR-Cas9 editing of exon 3 of the Tnfrsf14 locus. Red letters indicate mutated nucleotides. Green letters indicate protospacer adjacent motif (PAM) sequence. Blue letters indicate mutated amino acids. Black box shows restriction enzyme sites. (B) HVEM surface expression level of splenic CD4+ T cells, ILCs (CD3−Lin−CD90.2+), and iNKT cells (TCRβ+, CD1d tetramer+) from the indicated mice were determined by flow cytometry. HVEM-knockout mouse is a negative control for HVEM staining. KI, KI allele. ssDNA, single-stranded DNA.

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