Fig. 5: Selective recruitment of AGPATs to SARS-CoV-2 induced DMVs.

a Change of subcellular localization of AGPATs upon expression of SARS-CoV-2 nsp3-4. Huh7-derived cells transiently expressing AGPAT2-EGFP were transfected with a SARS-CoV-2 HA-nsp3-4-V5 expression construct or the empty vector. After 48 h, cells were stained with HA-specific antibody and examined by confocal microscopy. Maximum intensity projections are shown. Enrichment score indicates the likelihood of cells showing a punctate or diffuse staining pattern. Two biologically independent experiments showed similar results. b Clustering of AGPAT2-EGFP in SARS-CoV-2 HA-nsp3-4-V5 expressing cells. Huh7-Lunet/T7 cells were co-transfected with AGPAT2-EGFP and SARS-CoV-2 HA-nsp3-4-V5 or the empty vector. Twenty-four hours later, cells were fixed and ~1000 cells per condition were separated into two morphotypes (diffuse or punctate) using CellProfiler Analyst-based semi-supervised classifier. Significance was calculated using an ordinary one-way ANOVA. *p = 0.0211. A representative experiment from three biologically independent experiments is shown. c AGPAT clustering occurs independent of ER remodeling induced by nsp3-4. Huh7-Lunet cells expressing AGPAT2-EGFP and HA-nsp3-4-V5 were stained for the ER markers protein disulfide isomerase (PDI) and calnexin and analyzed by confocal microscopy. Two biologically independent experiments showed similar results. Light microscopy images are maximum intensity projections.