mouse anti hsv 1 vp16 monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Journal of Advanced Research

Article Title: Antimycin A inhibits alpha-herpesvirus replication by disrupting the formation of pyrimidinosomes

doi: 10.1016/j.jare.2025.05.016

Figure Lengend Snippet: Antimycin A exhibits extensive antiviral activity against alpha-herpesvirus. Antimycin A effectively inhibited HSV-1 (A-C) and HSV-2 (G-H) infections in Vero E6 cells, PRV (D-E) infection in PK-15 cells, and EHV-1 (J-K) infection in RK13 cells. Vero-E6 cells, PK-15 cells, and RK13 cells were pretreated for 12 h with increasing concentrations of Antimycin A and then infected with HSV-1 (A-C), PRV (D-E), HSV-2 (G-I), and EHV-1 (J-K) at MOIs of 0.5, 0.1, 0.5, and 0.5, respectively. At 24 hpi, cells were fixed and analyzed by fluorescence imaging. (A, D, G and J) Infection levels were quantified using a fluorescent microplate reader (black curve), while cell viability was measured using the CCK-8 Assay (orange curve). The CC50 for each compound was calculated via a four-parameter logistic nonlinear regression model in GraphPad Prism. Dotted lines indicate 50 % inhibition. Data represent the means ± SEM from n = 3 independent experiments of infectious virions, normalized to DMSO-treated wells. The IC50 values for HSV-1, PRV, HSV-2, and EHV-1 were determined by nonlinear regression analysis. (B, E, H and K) eGFP expression in infected cells, either untreated (0 μM) or treated with various concentrations (0.0015–5 μM) of Antimycin A, was visualized by fluorescence microscopy at the same time point. Representative images are shown. Bars, 300 µm. Magnification, ×10. (C, F and I) Western blot analysis was performed to quantify infection in cells infected with HSV-1, PRV, or HSV-2. For HSV-1, infection was assessed using ICP4, VP16, and gD as markers. For PRV, infection levels were quantified by immunoblotting for UL54. For HSV-2, infection was quantified by immunoblotting for VP16. β-actin was used as the loading control. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Primary antibodies used in this study included goat BHV-1 antisera, mouse anti-PRV UL47 monoclonal antibody (MAb), mouse anti-HSV-1 ICP4 monoclonal antibody (Santa Cruz, sc69809), mouse anti-HSV-1 VP16 monoclonal antibody (Santa Cruz, sc7545), mouse anti-HSV-1 gD monoclonal antibody (Santa Cruz, sc21719), and mouse anti-β-actin MAb (Proteintech, 66009–1-lg).

Techniques: Activity Assay, Infection, Fluorescence, Imaging, CCK-8 Assay, Inhibition, Expressing, Microscopy, Western Blot, Control

Journal: Journal of Advanced Research

Article Title: Antimycin A inhibits alpha-herpesvirus replication by disrupting the formation of pyrimidinosomes

doi: 10.1016/j.jare.2025.05.016

Figure Lengend Snippet: Antimycin A exhibits extensive antiviral activity against alpha-herpesvirus. Antimycin A effectively inhibited HSV-1 (A-C) and HSV-2 (G-H) infections in Vero E6 cells, PRV (D-E) infection in PK-15 cells, and EHV-1 (J-K) infection in RK13 cells. Vero-E6 cells, PK-15 cells, and RK13 cells were pretreated for 12 h with increasing concentrations of Antimycin A and then infected with HSV-1 (A-C), PRV (D-E), HSV-2 (G-I), and EHV-1 (J-K) at MOIs of 0.5, 0.1, 0.5, and 0.5, respectively. At 24 hpi, cells were fixed and analyzed by fluorescence imaging. (A, D, G and J) Infection levels were quantified using a fluorescent microplate reader (black curve), while cell viability was measured using the CCK-8 Assay (orange curve). The CC50 for each compound was calculated via a four-parameter logistic nonlinear regression model in GraphPad Prism. Dotted lines indicate 50 % inhibition. Data represent the means ± SEM from n = 3 independent experiments of infectious virions, normalized to DMSO-treated wells. The IC50 values for HSV-1, PRV, HSV-2, and EHV-1 were determined by nonlinear regression analysis. (B, E, H and K) eGFP expression in infected cells, either untreated (0 μM) or treated with various concentrations (0.0015–5 μM) of Antimycin A, was visualized by fluorescence microscopy at the same time point. Representative images are shown. Bars, 300 µm. Magnification, ×10. (C, F and I) Western blot analysis was performed to quantify infection in cells infected with HSV-1, PRV, or HSV-2. For HSV-1, infection was assessed using ICP4, VP16, and gD as markers. For PRV, infection levels were quantified by immunoblotting for UL54. For HSV-2, infection was quantified by immunoblotting for VP16. β-actin was used as the loading control. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Primary antibodies used in this study included goat BHV-1 antisera, mouse anti-PRV UL47 monoclonal antibody (MAb), mouse anti-HSV-1 ICP4 monoclonal antibody (Santa Cruz, sc69809), mouse anti-HSV-1 VP16 monoclonal antibody (Santa Cruz, sc7545), mouse anti-HSV-1 gD monoclonal antibody (Santa Cruz, sc21719), and mouse anti-β-actin MAb (Proteintech, 66009–1-lg).

Techniques: Activity Assay, Infection, Fluorescence, Imaging, CCK-8 Assay, Inhibition, Expressing, Microscopy, Western Blot, Control

Journal: eLife

Article Title: A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1

doi: 10.7554/elife.85412

Figure Lengend Snippet: Figure 4. ICP0 is a sufficient but not necessary condition of the ‘Escaping’. (A–E) dCas9-emerin cells infected with HSV-1 (2 hr on ice) at an MOI of 1 were electroporated with HSV-1 sgRNA or ctrl sgRNA at 1 hpi. (A) The mRNA levels of ICP27, TK, ICP0 and VP16 at the indicated time points were measured by qPCR. (B) The protein levels of ICP0, ICP8, TK and VP16 at the indicated time points were measured by immunoblotting. Data is shown as mean ± SD, n = 3. (C) dCas9-emerin cells were pretreated with PAA (500 µg/mL) or DMSO for 2 hr before infection and throughout the experiment and

Article Snippet: DOI: https://doi.org/10.7554/eLife.85412 19 of 25 (Abcam, #ab20194), mouse monoclonal anti- VP16 antibody (Santa Cruz, #sc- 7545), mouse monoclonal anti-β-actin antibody (Sino Biological, #1000166), anti- TK antibody (laboratory stock), rabbit anti- E- cadherin antibody (Affinity Biosciences, #AF0131), mouse monoclonal anti- GAPDH antibody (Abways, #AB0037), mouse monoclonal anti- Histone antibody (Sino Biological, #100005), rabbit polyclonal anti- dCas9 antibody (ABclonal, #A14997), goat anti- mouse IgG- HRP (Invitrogen, #31430), and goat anti- rabbit IgG (H+L)- HRP (Invitrogen, #32460).

Techniques: Infection, Western Blot