2 3 cgamp control  (InvivoGen)

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: Both oncogenes are needed to impact cGAMP levels. (A) western blot analysis for functional markers p53 and PTPN14, N= 3 using TTL system (B) cGAMP ELISA following DNA stimulation, N=4. (C) p-values for cGAMP ELISA of each cell line relative to HPV18 at specified time points. (D) Western blot showing function expression of E6 and/or E7 in overexpression cell lines. N=3 (E) cGAMP ELISA following DNA transfection, N=3 Statistics for (C and E) were calculated with two-way ANOVA, followed by Dunnett’s multiple comparison test. * p < 0.05; **p < 0.01; *** p < 0.001. Error bars indicate SEM.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Western Blot, Functional Assay, Enzyme-linked Immunosorbent Assay, Expressing, Over Expression, Transfection, Comparison

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: HPV18(+) cells produce more cGAMP than parental control cells. Quantification of cGAMP by ELISA following DNA stimulation of HPV(-) cells (black) and HPV18(+) cells (red). N=3 donors, 3 reps in each donor, average of the 9 replicates, (A) replicates separated by donor (B). Statistics for (A) were calculated using a linear mixed-effects model with “HPV” and “Time” as fixed effects and donor as a random effect to account for inter-donor variability. Statistics for (B) were calculated with a two-way ANOVA followed by a Dunnett’s post hoc test. * p<0.05; ** p<0.01; *** p<0.001Error bars for represent SEM, calculated by first averaging technical replicates (n=3 per donor) with propagated error, and then computing SEM across biological replicates (n=3 donors).

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: HPV decreases STING and IRF3 activation. (A) Western blot showing DNA-treated HPV18(+) and parental cells. Quantification of (B) pSTING, average of the 9 replicates (C) pSTING, separated by donor (D) pIRF3 western blots, average of the 9 replicates, (E) pIRF3, separated by donor. N=3 donors, 3 replicates in each. Statistics for the average of 9 (B and D) were calculated using a linear mixed-effects model with “HPV” and “Time” as fixed effects and donor as a random effect to account for inter-donor variability. Statistics for each cell line (C and E) were calculated with a two-way ANOVA followed by a Dunnett’s post hoc test. * p < 0.05; ** p<0.01; *** p<0.001. Error bars represent SEM, calculated by first averaging technical replicates (n=3 per donor) with propagated error, and then computing SEM across biological replicates (n=3 donors). Slope was calculated by fitting linear regressions within each time interval, and the values were subsequently analyzed using a linear mixed-effects model.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Activation Assay, Western Blot

Journal: bioRxiv

Article Title: Human Papillomavirus Infection Blunts STING Signaling and Alters Downstream Response

doi: 10.1101/2025.11.06.686957

Figure Lengend Snippet: Both oncogenes are needed to impact STING activation. (A) Western blot for total STING, pSTING, and αTubulin for the TTL cell lines, N=3. (B) Densitometry of pSTING normalized to αTubulin and a loading control lysate run on all blots. (C) p-values for pSTING densitometry of each cell line relative to HPV18 cells at specified time points. (D) Western blot for total STING, pSTING, and αTubulin for the over expression cell lines, N=3. (E) Densitometry of pSTING normalized to αTubulin and a loading control lysate run on all blots. (F) p-values for pSTING densitometry of each cell line relative to E6/E7 expressing cells at specified time points. Statistics for (C and F) were calculated with two-way ANOVA, followed by Dunnett’s multiple comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001. Error bars indicate SEM.

Article Snippet: Primary human foreskin keratinocytes (HFKs) with or without HPV18 genomes were treated with 25 μg/mL 2′3′-cGAMP (InvivoGen tlrl-nacga23) to directly activate STING.

Techniques: Activation Assay, Western Blot, Control, Over Expression, Expressing, Comparison

Journal: Vaccines

Article Title: Current Status of Porcine Reproductive and Respiratory Syndrome Vaccines

doi: 10.3390/vaccines12121387

Figure Lengend Snippet: Key challenges for PRRS vaccines and some possible solutions.

Article Snippet: N, M, GP5 and E proteins , Baculovirus/Sf9 cells , Entrapped in VLP nanoparticles , 3 weeks , 2′,3′-cGAMP VacciGradeTM (Invivogen, USA) , IN , Enhanced viremia associated with IFN-α, IFN-γ, and IL-10. , [ ] .

Techniques: Vaccines, Virus, Modification, Sequencing, Adjuvant, Infection, Selection, Control, Injection, Activity Assay, Multiplex Assay

Journal: Vaccines

Article Title: Current Status of Porcine Reproductive and Respiratory Syndrome Vaccines

doi: 10.3390/vaccines12121387

Figure Lengend Snippet: Some explorations of improving PRRSV inactivated vaccine-induced immune efficacy.

Article Snippet: N, M, GP5 and E proteins , Baculovirus/Sf9 cells , Entrapped in VLP nanoparticles , 3 weeks , 2′,3′-cGAMP VacciGradeTM (Invivogen, USA) , IN , Enhanced viremia associated with IFN-α, IFN-γ, and IL-10. , [ ] .

Techniques: Virus, Emulsion, Infection, Purification, Modification, Expressing, Activation Assay, Immunopeptidomics

Journal: Vaccines

Article Title: Current Status of Porcine Reproductive and Respiratory Syndrome Vaccines

doi: 10.3390/vaccines12121387

Figure Lengend Snippet: The studies of PRRSV subunit vaccines.

Article Snippet: N, M, GP5 and E proteins , Baculovirus/Sf9 cells , Entrapped in VLP nanoparticles , 3 weeks , 2′,3′-cGAMP VacciGradeTM (Invivogen, USA) , IN , Enhanced viremia associated with IFN-α, IFN-γ, and IL-10. , [ ] .

Techniques: Vaccines, Expressing, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Transformation Assay, Virus, Modification, Adjuvant, Synthesized, Infection