Fig. 8: AIF participates in LPS/Ng-induced cell death.

A THP-1 cells (shNC and shTFGs) were stimulated with LPS (1 μg/mL) for 4 h followed by Nigericin (5 μg/mL) stimulation for 60 min (LPS/Ng). Z-VAD plus NEC-1 were used to block apoptosis or necrosis. Cell death were determined using SYTOX staining. B Scatter dot plots of cell death percentage in groups described in A are shown. Data are shown as mean ± s.d. n.s. no significance; n = 3 for each group. C THP-1 cells were transduced with TFG-specific shRNAs. Cells were stimulated with LPS/Ng. Nuclear/Cytosol fractions were extracted and IB assays were performed to analyze AIF localization. D THP-1 cells were transduced with shNC, shTFG, or shULK1. Cells were stimulated with LPS/Ng. Mitochondrial/non-mitochondrial fractions were extracted and IB assays were performed to analyze AIF localization. COX4 was detected as a mitochondrial marker. E THP-1 cells were transduced with shNC or shTFG. TFG stable knockdown THP-1 cells were then transfected with LV8N-(mcherry)-ULK1 to rescue ULK1 expression (shTFG + ULK1res). Mitochondrial/non-mitochondrial fractions were extracted and IB assays were performed to analyze AIF localization. COX4 was detected as a mitochondrial marker. Experiments were independently repeated three times.