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. 2018 Sep 20;3(18):e120179. doi: 10.1172/jci.insight.120179

Figure 7. IFI16•dsDNA filaments are released from epithelial cells following exposure to cytotoxic lymphocyte granule contents.

Figure 7

(A and B) IFN-treated human salivary gland (HSG) cells were transfected with biotinylated DNA to induce IFI16 filament formation and were then mock treated (A) or exposed to YT cell granule contents (GC) for 3 hours (B). Cells were fixed and stained with DAPI (blue), anti-IFI16 antibody (green), and Streptavidin-DyLight 594 (red). (C) Induction of apoptosis by GC treatment was confirmed by Western blotting for caspase 3, demonstrating intact (closed arrow) and cleaved (open arrow) forms. (D) Following GC treatment, samples of cell lysate (top panel) and supernatant (middle panel) were collected and analyzed in parallel for IFI16 and DNA content by blotting with anti-IFI16 antibody and StrepTactin-HRP. Biotinylated DNA was isolated from the supernatant of treated cells using Streptavidin DynaBeads, and the presence of IFI16•dsDNA interaction in the supernatant was confirmed by blotting for each (D, bottom panel). Scale bars: 10 μM. Data are representative of results of 3 separate experiments.