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. 2023 May 25;186(11):2380–2391.e9. doi: 10.1016/j.cell.2023.04.024

Figure S1.

Figure S1

Comparison of EABR-related sequence insertions in the cytoplasmic tail of SARS-CoV-2 S, related to Figure 1

(A) Top: schematic of different S-EABR constructs that were compared for their ability to induce eVLP assembly. EPM, endocytosis prevention motif; GS, (Gly)3Ser linker; EABR, ESCRT- and ALIX-binding region. Bottom: amino acid sequences of EABR portion of different constructs.

(B) Western blot analysis of SARS-CoV-2 S1 protein levels on eVLPs purified by ultracentrifugation on a 20% sucrose cushion from transfected Expi293F cell culture supernatants. Cells were transfected with S-p6, S-VP401–44, S-p9, or S-EABR constructs. Purified eVLP samples were diluted 1:400.

(C) Western blot analysis comparing HIV-1 EnvYU2 levels in eVLP samples purified from transfected Expi293F cell culture supernatants. Cells were transfected with plasmids encoding Env-EABR, Env plus HIV-1 Gag, or Env alone. Purified eVLP samples were diluted 1:200.

(D) Western blot analysis comparing CCR5 levels in eVLP samples purified from transfected Expi293F cell culture supernatants. Cells were transfected with plasmids encoding CCR5-EABR, CCR5 plus HIV-1 Gag, or CCR5 alone. Purified eVLP samples were diluted 1:200. The migration difference between CCR5-EABR and CCR5 is due to addition of the EABR sequence (∼7 kDa) that increases its molecular mass.