Figure 2.

STING1 interacts with the components of the MTORC1 complex. (A) Coimmunoprecipitation and immunoassay of extracts of HEK293T cells transfected with various combinations of expression vector for MYC-STING1 and other indicated plasmids. (B) Coimmunoprecipitation and immunoassay of extracts of Hep3B cells transfected with Flag-RPTOR, MYC-STING1, and transfected with cGAMP (2 μg/ml, 6 h), HT-DNA (10 μg/ml, 6 h), or treated with PA (0.5 mM, 24 h). (C-E) Confocal microscopy of Hep3B cells (C and E), or mouse primary hepatocytes with or without mouse-Sting1-siRNA (MmSting1-siRNA) transfection (D) were treated with PA (0.5 mM, 24 h) or cGAMP (2 μg/ml, 6 h). Cells treated with scrambled siRNA were used as control. The cells were fixed 8 h later and stained with indicated antibodies. Hoechst 33342 was used for nucleus staining. Representative images are shown. Scale bar: 10 μm. For (E), the right panels of enlarged images correspond to rectangular frames in the left single or overlapped results from the same group. For the overlapped results, the enlarged images are listed in the following order: the left-upper image in the enlarged part is associated with the overlapped STING1-LAMP1; the left-lower image is associated with the overlapped STING1-RPTOR; the right-upper image is associated with the overlapped LAMP1-RPTOR; the right-lower image is associated with the overlapped STING-LAMP1-RPTOR. (F) Coimmunoprecipitation and immunoassay of extracts of Hep3B cells transfected with HA-RPTOR, Flag-RRAGA, and transfected with cGAMP (2 μg/ml, 6 h), or treated with PA (0.5 mM, 24 h). (G) Coimmunoprecipitation and immunoassay of extracts of Hep3B cells transfected with MYC-MLST8, Flag-RRAGA, and transfected with cGAMP (2 μg/ml, 6 h), or treated with PA (0.5 mM, 24 h).