Supplementary Figure 12: Presentation of mtDNA neoantigens via MHC.
From: De novo mutations in mitochondrial DNA of iPSCs produce immunogenic neoepitopes in mice and humans
a, iPSCs (P7) underwent CRISPR-Cas9 inactivation of the B2m and Ciita genes to generate double MHC knockout (dKO) iPSCs and iEC (dKO) were differentiated. b, The knockouts of B2m and Ciita in iEC (dKO) were confirmed by PCR (representative gel of two independent experiments). c, Surface expression of MHC class I and II was assessed by flow cytometry (mean ± s.d., 4 independent experiments per group, two-tailed Student’s t-test). d, B/6 mice were immunized with either iEC (P7) or iEC (dKO) overexpressing B/c (red) or B6 (blue) Co3 and Cytb. The splenocyte response after 5 days against the same overexpressed proteins in either iEC (P7) or iEC (dKO) was assessed in Elispot assays to determine the mechanistic role of MHC. e, Elispot assays of B/6 mice immunized with iEC (P7) or iEC (dKO) overexpressing syngeneic B/6 proteins are shown (mean ± s.d., quadruplicates of 3 animals per group, two-tailed Student’s t-test). f, Elispot assays of B/6 mice immunized with iEC (P7) or iEC (dKO) overexpressing allogeneic B/c proteins are shown. Unstimulated responder splenocytes served a control (mean ± s.d., quadruplicates of 3 animals per group, two-tailed Student’s t-test).
