Journal: Frontiers in Pharmacology

Article Title: Evaluation and mechanism study of Pien Tze Huang against EV-A71 infection

doi: 10.3389/fphar.2023.1251731

Figure Lengend Snippet: PZH protects mice from EV-A71 infection. (A–C) 11-Day ICR mice were intraperitoneally inoculated with EV-A71-H-MA (20000 TCID 50 ) for 1 h. Following the infection, the infected mice received intragastric administration of PZH (170, 56.7 and 18.9 mg/kg) and intraperitoneal injection of RBV (50 mg/kg) daily for 7 days. The survival of the mice was monitored daily for 14 days (A, B) and the clinical scores were recorded for the same duration (C) . **p < 0.01 by A Log-Rank (Mantel-Cox) assay (A,B) and ***p < 0.001 by Ridit assay (C) . (D–F) The mice were dissected at day 5 post-infection. Muscle tissues were subjected to viral titer assays [ (D) , n = 5]. Immunohistochemistry analyses were performed on muscle tissue sections using EV-A71 VP1 antibody [ (E) , n = 5)]. Paraffin-embedded sections of muscle tissues were prepared and examined using H&E stain [ (F) , n = 5]. *p < 0.05, **p < 0.01 and ***p < 0.001 by one-way ANOVA with Holm-Sidak multiple comparisons test or Ridit assay, compared to the vehicle group. *p < 0.05, **p < 0.01 and ***p < 0.001 by one-way ANOVA with Holm-Sidak multiple comparisons test (D,E) or Ridit assay (F) , compared with vehicle group.

Article Snippet: The cells were incubated with GAPDH antibody (Proteintech) and EV-A71 VP1 antibody (GeneTex) at room temperature for 2 h and washed with PBS for three times.

Techniques: Infection, Injection, Immunohistochemistry, Staining

Journal: Frontiers in Pharmacology

Article Title: Evaluation and mechanism study of Pien Tze Huang against EV-A71 infection

doi: 10.3389/fphar.2023.1251731

Figure Lengend Snippet: Validation of differently expression proteins. RD cells were mock-infected or infected with EV-A71 (H, MOI = 0.01) for 1 h. Then, cells were subjected to treatment with either PZH or RBV for 24 h. The cells were harvested and the expression of VP1 (A) , p-AKT (B) , p-mTOR (C) , p-PI3K (D) , p-p65 (E) , p-JNK (F) was examined by Western blot. *p < 0.05, **p < 0.01 by t -test.

Article Snippet: The cells were incubated with GAPDH antibody (Proteintech) and EV-A71 VP1 antibody (GeneTex) at room temperature for 2 h and washed with PBS for three times.

Techniques: Expressing, Infection, Western Blot

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: HeLa cells were mock-infected and infected with EV-A71 (MOI 1) for 0, 12, and 24 h. PK-15 cells were mock-infected and infected with SVV (MOI 1) or FMDV (MOI 0.5) for 0, 6, and 12 h. The ΔΨm (A), mitochondrial Ca 2+ concentration (B), mitochondrial ROS (C), and mPTP opening (D) were detected using JC-1, Rhod-2 AM, mitoSOX, and calcein-AM, respectively. Data for CCCP (A) and lono (D) positive controls are shown. Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Infection, Concentration Assay, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: HeLa cells were mock-infected or infected with EV-A71 (MOI 1) for 0, 12, and 24 h, while PK-15 cells were mock-infected or infected with SVV (MOI 1) or FMDV (MOI 0.5) for 0, 6, and 12 h. The lysates were immunoprecipitated with anti-cGAS antibody. The mtDNA and gDNA in cGAS pulldown samples were detected and analyzed by qPCR. The EGFP DNA was used as an internal control (A). HeLa cells were mock-infected or infected with EV-A71 (MOI 1) for 24 h. PK-15 cells were mock-infected or infected with SVV (MOI 1) or FMDV (MOI 0.5) for 12 h. The mtDNA release was evaluated by IFA. Cells were double-immunostained for detection of Tom20 (red) and dsDNA (green); cellular nuclei were counterstained with 4’,6-diamidino-2-phenylindole (DAPI) (blue) (B, C, and D). Scale bar, 10 μm. Oropharyngeal tonsils collected from pigs infected with SVV (E) or FMDV (F) were analyzed for mtDNA release. Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Infection, Immunoprecipitation, Control, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: HT-29 WT and PPID -/- cells were mock-infected or infected with EV-A71 (MOI 1) for 24 h, while PK-15 WT or PPID -/- cells were mock-infected or infected with SVV (MOI 1) or FMDV (MOI 0.5) for 12 h. The mtDNA release was evaluated by qPCR (A). The expression of the IFN-β protein was detected by ELISA kit, and the expression of PPID was confirmed by Western blotting (B). (C) HT-29 cells were mock-infected or infected with EV-A71 (MOI 1), while PK-15 cells were mock-infected or infected with SVV (MOI 1) or FMDV (MOI 0.5). After 1 h of incubation of the virus or DMEM, cells were treated with DMSO or VBIT4 inhibitor (10 uM) for 24 or 12 h. The mtDNA release was then detected by qPCR. The expression of Bax protein in the WT and Bax -/- cells was confirmed by western blotting (D) and these cells were used to measure mtDNA release after infection with EV-A71 (HT-29 cells at MOI 1) or SVV (PK-15 cells at MOI 1) or FMDV (PK-15 cells at MOI 0.5) for 12 h (E). Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Infection, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Incubation, Virus, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: PK-15 cells were transfected with 2 μg of empty vector, the indicated SVV protein expressing plasmids (A) or FMDV protein expressing plasmids (B), and HeLa cells were transfected with 2 μg of empty vector or EV-A71 protein expressing plasmids (C). At 24 hpt, the mtDNA release was analyzed by qPCR. The expression of viral proteins was determined by Western blotting. (D) HeLa cells were transfected with 2 μg of empty vector, EMCV 2B or CA16 2B expressing plasmids for 24 h. The mtDNA release was detected by qPCR. (E) HT-29 WT and PPID -/- cells were transfected with 2 μg of empty vector, EMCV 2B, CA16 2B, or EV-A71 2B expressing plasmids for 24 h, and PK-15 WT or PPID -/- cells were transfected with 2 μg of empty vector, SVV 2B or FMDV 2B expressing plasmids for 24 h. The mtDNA release was detected by qPCR. Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Transfection, Plasmid Preparation, Expressing, Western Blot, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: IFN-β and ISG54 mRNA expression in cGAS -/- HeLa cells infected with SVV (A), and cGAS -/- HT-29 cells infected with EV-A71 (B); IFN-β protein expression is also shown (C). Western blotting was used to detect the expression of SVV VP1 protein (D) and EV-A71 3C protein (E) and viral titers were determined by TCID 50 assay. Bone marrow-derived macrophages from the WT or cGAS -/- mice were infected with FMDV and IFN-β protein in the supernatant was determined by ELISA (F). The expression of FMDV VP1 protein was detected by Western blotting, and the viral titers were determined by TCID 50 assay (G). WT and ρ 0 PK-15 cells were infected with SVV or FMDV and WT and ρ 0 HT-29 cells were infected with EV-A71. IFN-β protein expression level was determined by ELISA (H), and the viral titers were determined by TCID 50 assay (I). Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Expressing, Infection, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: (A) The 3-weeks-old WT and cGAS -/- mice were showed, and the expression of cGAS in the WT and cGAS -/- mice was detected by Western blotting. (B and C) The three-day-old WT mice were subcutaneously inoculated with FMDV (10 8 TCID 50 ) or EV-A71 (10 8 TCID 50 ) for 0, 1, or 3 d (n = 3/group), the mice carcasses without the head, tail, limbs, and viscera were collected for detection of mtDNA release (B) and IFN-β mRNA expression (C). (D and E) The three-day-old WT (n = 8) and cGAS -/- (n = 8) mice were subcutaneously inoculated with FMDV (10 8 TCID 50 ) or EV-A71 (10 8 TCID 50 ). The IFN-β mRNA level in the carcasses without the head, tail, limbs, and viscera from FMDV-infected mice was measured and compared at 3 dpi by qPCR (D). The IFN-β mRNA level in the carcasses without the head, tail, limbs, and viscera from EV-A71-infected mice was measured and compared at 1 dpi by qPCR (E). FMDV (F) and EV-A71 (G) titers in the mice carcasses without the head, tail, limbs, and viscera were determined at 2 dpi by TCID 50 assay. The mortality of WT (n = 10) and cGAS -/- (n = 10) mice infected by FMDV (F) or EV-A71 (G) was determined, respectively. Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Expressing, Western Blot, Infection, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: (A) HEK-293T cells were transfected with 1 μg of empty vector or the indicated FLAG-2C-expressing plasmids, along with 1 μg of empty vector or HA-cGAS plus HA-STING expressing plasmids. At 24 hpt, the IFN-β protein amount in the supernatant was determined by ELISA, and the expression of 2C was confirmed by Western blotting. (B) STING-HEK-293T cells were transfected with increasing amount (0, 3, or 6 μg) of the indicated FLAG-2C-expressing plasmids. At 24 hpt, cells were transfected with poly(dA:dT) (2 μg/ml) for 12 h. The cells lysates were immunoprecipitated with anti-STING antibody. The antibody-antigen complexes were detected using anti-STING, TBK1, and FLAG antibodies, respectively. (C) Schematic representation of the structure and conserved functional sites in EV-A71 2C protein. The redder the color was, the more conservative the sites were. (D, E) HEK-293T cells were transfected with 1 μg of empty vector or EV-A71 (D), CA16 (E), and EMCV (E) FLAG-2C- or FLAG-2C mutants-expressing plasmids, along with 1 μg of empty vector or HA-cGAS plus HA-STING expressing plasmids. At 24 hpt, the IFN-β protein amount in the supernatant was determined by ELISA, and the expression of 2C was confirmed by Western blotting. (F) STING-HEK-293T cells were transfected with 6 μg of empty vector, FLAG-2C- or FLAG-2C mutants-expressing plasmids. At 24 hpt, cells were transfected with poly(dA:dT) (2 μg/ml) for 12 h. The cells lysates were immunoprecipitated with anti-STING antibody. The antibody-antigen complexes were detected using anti-STING, TBK1, and FLAG antibodies, respectively. (G) HEK-293T cells were transfected with 1 μg of empty vector, FLAG-2C- or FLAG-2C mutants-expressing plasmids, and 1 μg of HA-cGAS plus HA-STING expressing plasmids. At 24 hpt, expression of IRF3, p-IRF3, and FLAG-2C protein was determined by Western blotting. The IRF3 dimerization was detected using native PAGE. Error bars show standard deviation. **, P <0.01.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Transfection, Plasmid Preparation, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Immunoprecipitation, Functional Assay, Clear Native PAGE, Standard Deviation

Journal: PLOS Pathogens

Article Title: Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release

doi: 10.1371/journal.ppat.1011132

Figure Lengend Snippet: In this model, cytosolic mtDNA released after EV-A71, SVV, and FMDV infection binds to cGAS to activate cGAS-mediated signal transduction, resulting in the antiviral responses. Virus immune evasion strategies include SVV 2C induced the reduction of cGAS by activation of autophagy and FMDV 2B and 3C pro proteins which inhibit STING expression to block the antiviral response. Furthermore, EV-A71, CA16, and EMCV 2C can antagonize activation of the cGAS-STING signaling pathway by impairing the interaction of STING with TBK1, while FMDV L pro and EV-A71 2A proteins inhibit cGAS-STING-induced IFN-β protein expression.

Article Snippet: The commercial antibodies used in this study included an anti-FLAG monoclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), anti-FLAG polyclonal antibody (Sigma-Aldrich), anti-HA monoclonal antibody (Thermo Scientific, Waltham, MA, USA), anti-cGAS monoclonal antibody (Santa Cruz Biotechnology), anti-STING monoclonal antibody (Cell Signaling Technology, Beverly, MA, USA), anti-p-STING (S366) monoclonal antibody (Cell Signaling Technology), anti-IFI16 monoclonal antibody (Cell Signaling Technology), anti-TBK1 monoclonal antibody (Cell Signaling Technology), anti-IRF3 monoclonal antibody (Cell Signaling Technology), anti-p-IRF3 monoclonal antibody (Cell Signaling Technology), anti-dsDNA monoclonal antibody (Abcam, Cambridge, MA, USA), anti-ATG7 polyclonal antibody (ABclonal Technology Co., Ltd, Wuhan, China), anti-Tom20 monoclonal antibody (ABclonal), and anti-β-actin monoclonal antibody (Thermo Scientific), anti-Bax monoclonal antibody (ABclonal), anti-PPID polyclonal antibody (ABclonal), anti-EV-A71 3C polyclonal antibody (ABclonal), anti-SVV VP1 polyclonal antibody was prepared in our laboratory [ ], anti-FMDV VP1 polyclonal antibody was prepared in our laboratory [ ], and anti-2C polyclonal antibody was produced in rabbits by immunization with SVV 2C protein.

Techniques: Infection, Transduction, Virus, Activation Assay, Expressing, Blocking Assay