Fig. 6
Interaction of the N-terminal region of SP110 with SP110b and NF-κB proteins. a HEK293T cells were co-transfected with the respective constructs, and whole-cell HEK293T extracts were subjected to immunoprecipitation (IP) using an anti-FLAG antibody and analyzed by WB with the indicated antibodies at 2 days post-transfection. b and c HEK293T cells were co-transfected with the indicated constructs, and the cellular distribution of eGFP-SP110 fusion proteins that contain deletion mutants of SP110 protein (b and c), FLAG-SP110b (b) and FLAG-p50 (c) was visualized using confocal microscopy at 2 days post-transfection. Scale bars: 10 μm. The cells were probed with an anti-FLAG antibody to identify FLAG-SP110b (b) and FLAG-p50 (c), respectively, and subjected to Hoechst staining to identify nuclei. d HEK293T cells were co-transfected with the constructs expressing eGFP-SP1101–276 and FLAG-p50, and the cells were probed with an anti-FLAG antibody to identify FLAG-p50 and visualized using microscopy at 2 days post-transfection. The numbers of cells with p50 cytoplasmic or nuclear distribution were determined by counting at least 60 cells per field from two randomly selected microscopic fields. Mean number of cells with p50 in cytoplasm or nucleus was shown. The data are presented as the mean ± SD. Statistical significance of the difference between two sample groups was calculated using a two-tailed unpaired t-test. **P < 0.01. e HEK293T cells were co-transfected with the respective constructs, and whole-cell HEK293T extracts were subjected to IP using an anti-FLAG or control IgG antibody and analyzed by WB with the indicated antibodies at 2 days post-transfection. All data represent at least 2 independent experiments