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. 2021 Aug 12;18(4):860–876. doi: 10.1080/15548627.2021.1961072

Figure 1.

Figure 1.

PA treatment stimulates MTORC1 activity through STING1. (A) Hep3B cells were transfected with 2ʹ3’-cGAMP for indicated concentration. After 6 h, lysates were immunoblotted for p-RPS6KB1 (T389), RPS6KB1, p-RELA (S536), RELA, p-TBK1 (S172), TBK1, and ACTB. (B) Lysates of Hep3B cells transfected with the indicated concentration of HT-DNA for 12 h, or treated with the indicated concentration of PA for 24 h, were used for immunoblotting with the p-RPS6KB1 (T389), RPS6KB1, p-TBK1 (S172), TBK1, p-RPS6 (S235/236), RPS6, and ACTB antibodies. (C) Control (GFP-KO) and STING1-KO Hep3B cells were treated with PA (24 h) for indicated concentration, and lysates were immunoblotted for p-RPS6KB1 (T389), RPS6KB1, p-RPS6 (S235/236), RPS6, STING1, and ACTB. (D) Immunoassay of extracts of control (GFP-KO) or STING1-KO Hep3B cells treated with 2ʹ3’-cGAMP (cGAMP, 2 μg/ml) for the indicated time interval. (E) Hep3B cells with or without STING1-siRNA (STING1-siRNA) transfection were treated with HT-DNA (10 μg/ml) for the indicated time interval were used for immunoblotting with the indicated antibodies. Cells treated with scrambled siRNA were used as control. (F) Mouse primary hepatocytes with or without mouse-Sting1-siRNA (MmSting1-siRNA) transfection were treated with PA (0.5 mM, 24 h), or transfected with cGAMP (2 μg/ml, 6 h), and the lysates of cells were performed for immunoblotting. Cells treated with scrambled siRNA were used as control.