Fig. 2

TRAF3 knockout (TRAF3KO) cells activate type-I interferon (IFN)-related pathways. A Western blot of TRAF3KO clones demonstrating the depletion of TRAF3 compared to the wild-type (WT) TRAF3 of ITB1 lysates. B Growth of ITB1 and TRAF3KO cells in culture was monitored every 8 h until confluency. The cyan line represents the growth of ITB1 cells, and the pink lines represent the growth of TRAF3KO cells. C Quantitative PCR shows the mRNA levels of genes related to the type-I IFN pathway (IFNA, IFNB, IRF7, IFIT1, ISG15). Cyan and pink bars represent the levels in ITB1 and TRAF3KO cells, respectively. D Western blot of proteins in the STING pathway in ITB1 and TRAF3KO lysates. Actin and histone H3 were used as loading controls. Numbers in parentheses represent the protein size in kDa. E Immunofluorescence images showing the expression of STING (Cy3) in the nucleus (DAPI) of ITB1 and TRAF3KO cells. F Flow cytometry results showing the surface levels of MHC-I using an antibody conjugated to the FITC fluorophore. The mean fluorescence intensities (MFI) were compared between each condition and plotted on a bar graph. G Kaplan–Meier plot showing the survival of mice in four groups: NSG mice injected with ITB1 (solid pink line), NSG mice injected with TRAF3KO cells (dotted pink line), WT mice injected with ITB1 (solid cyan line), and WT mice injected with TRAF3KO cells (dotted cyan line)