Journal: bioRxiv

Article Title: Proximity proteomics reveals a role for IFI16 during human coronavirus infection

doi: 10.64898/2026.04.22.720112

Figure Lengend Snippet: IFI16 promotes coronavirus infection. ( a) CRISPR-Cas9 technology was used to knock out IFI16 in A549 cells and single cell clones were screened for loss of IFI16 expression using qRT-PCR. Data is representative of two independent experiments. ( b) Immunoblot showing the loss of IFI16 expression in IFI16-KO cell line. ( c) Immunofluorescence assay showing reduced number of HCoV-OC43 N-positive cells upon loss of IFI16. Cells were infected at an MOI of 0.1 and incubated for the indicated length of time before fixation and staining for HCoV-OC43 N protein. Scale bar is 10 um. ( d ) Representative flow cytometry analysis of wildtype (WT) A549 cells and IFI16-KO cells infected (in quadruplicate) with HCoV-OC43 at an MOI of 0.01 and 0.1 for 12, 24 and 48 h. Data is representative of 3 independent experiments. (e) and (f) Quantification of flow cytometry data in . ( g) Representative flow cytometry analysis of CRISPRi cell lines infected (in quadruplicate) with HCoV-OC43 at an MOI of 0.01 for 12, 24 and 48 h. data in figure. (h) Quantification of flow cytometry data in . Data is representative of two independent experiments. Statistical significance was assessed by ANOVA. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001.

Article Snippet: Cells were washed twice with a permeabilization/wash buffer (0.1% saponin and 0.5% BSA in 1X PBS) and stained with a 1:10000 dilution of anti-HCoV-OC43-N rabbit polyclonal antibody (catalogue no. 40643-T62; Sino Biological) for 30 min at 4°C.

Techniques: Infection, CRISPR, Knock-Out, Single Cell, Clone Assay, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Incubation, Staining, Flow Cytometry