Fig. 4
NRF2 interacts with NICD and mediates STING activation in macrophages. Bone marrow-derived macrophages (BMMs, 1 × 106) were isolated from PTENM−KO and PTENFL/FL mice and then stimulated with LPS (100 ng/ml) for 6 h. A Immunofluorescence staining of NICD (red) in BMMs from PTENM−KO and PTENFL/FL after LPS stimulation. Scale bars, 20 μm. Data representative of three experiments. B Immunofluorescence staining of NRF2 (green) in BMMs from PTENM−KO and PTENFL/FL after LPS stimulation. DAPI was used to visualize nuclei (blue). Scale bars, 20 μm. Data representative of three experiments. C Immunofluorescence staining for macrophage NICD (red) and NRF2 (green) in LPS-stimulated macrophages. Scale bars, 20 μm. Data representative of three experiments. D Western-blot analysis and relative density ratio of NICD and NRF2 in the PTENFL/FL and PTENM−KO macrophage nucleus after LPS stimulation (n = 6 samples/group). E Immunoprecipitation analysis of NRF2 and NICD in LPS-stimulated macrophages from PTENM−KO mice. Data representative of three experiments. F BMMs were transfected with the CRISPR-Notch1 KO vector (1, 2, and 3ug) or control. Immunoprecipitation analysis of NRF2 and RBPjκ in LPS-stimulated macrophages from PTENM−KO mice. Data representative of three experiments. G Immunoprecipitation analysis of NRF2 and RBPjκ in LPS-stimulated macrophages from PTENFL/FL and PTENM−KO mice. Data representative of three experiments. H BMMs were transfected with the CRISPR-Notch1 KO vector or control. Western blot analysis and relative density ratio of NICD, NRF2, p-STING, STING, p-TBK1, TBK1, p-P65, and P65 in PTENM−KO BMMs transfected with the CRISPR-Notch1 KO vector or control after LPS stimulation (n = 6 samples/group). All data represent the mean ± SD. ***p < 0.001, ****p < 0.0001