Fig. 6
STING plays a critical role in the PTEN‑mediated inflammatory response and ROS generation in LPS‑stimulated macrophages. Bone marrow-derived macrophages (BMMs) were isolated from PTENM−KO or PTENFL/FL mice and then transfected with CRISPR/Cas9-mediated STING activation (ACT) or STING knockout (KO) vector. A Western blot analysis and relative density ratio of STING, p-TBK1, TBK1, p-P65, and P65 in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING ACT and control from PTENM−KO mice (n = 6 samples/group). B Quantitative RT-PCR analysis of Ifn-β, Tnf-α, IL-1β, Cxcl-1, and Mcp-1 mRNA levels in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING ACT and control from PTENM−KO mice (n = 6 samples/group). (C) Detection of ROS generation by Carboxy-H2DFFDA in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING ACT and control from PTENM−KO mice. Quantification of ROS-producing macrophages (green) (n = 6 samples/group). Scale bars, 100 μm. D Quantitative RT-PCR analysis of Ifn-β, Tnf-α, IL-1β, Cxcl-1, and Mcp-1 mRNA levels in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING KO and control from PTENFL/FL mice (n = 6 samples/group). E Western blot analysis and relative density ratio of STING, p-TBK1, TBK1, p-P65, and P65 in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING KO and control from PTENFL/FL mice (n = 6 samples/group). F Detection of ROS generation by Carboxy-H2DFFDA in LPS-stimulated BMMs transfected with CRISPR/Cas9-mediated STING KO and control from PTENFL/FL mice. Quantification of ROS-producing macrophages (green) (n = 6 samples/group). Scale bars, 100 μm. All data represent the mean ± SD. **p < 0.01, ***p < 0.001, ****p < 0.0001