Journal: Cell Insight

Article Title: IRF1 amplifies HSV-1-triggered antiviral innate immunity in a feed-forward manner

doi: 10.1016/j.cellin.2025.100255

Figure Lengend Snippet: Fig. 3. IRF1 restricts HSV-1 replication. (A) HT1080 cells transduced with control sgRNA (Ctrl) or sgRNA targeting IRF1 were infected with HSV-1 (MOI = 0.01). Viral titers in the supernatants were quantified at 48 h post-infection. (B–E) THP-1 cells transduced with control sgRNA (Ctrl) or sgRNA targeting IRF1 were mock- infected or infected with HSV-1 (MOI = 1). The expression of ICP0, ICP8, and UL19 was quantified by RT-qPCR (B–D). The mock-infected samples were labeled as N. D. (not detected), and the signals from wild-type (WT) cells infected with HSV-1 were normalized to 1. WCLs were analyzed by immunoblotting at 8 h post-infection (E). (F) HEK293T cells were transfected with an IFN-β promoter reporter plasmid mixture with increasing amounts of IRF1 expression plasmids (0, 0.1, 0.2, or 0.5 μg). Luciferase activities were measured at 24 h post-transfection. (G–H) HT1080 cells stably expressing vector control, IRF1-WT, or IRF1-R82A were infected with HSV- 1-GFP (MOI = 0.05), and GFP expression was imaged at 24 h post-infection (G). Scale bars,100 μm. Viral titers in the supernatants were quantified at 24 h post- infection (H).

Article Snippet: The following antibodies and reagents were used for immunoblotting and immunoprecipitation: Mouse anti-FLAG monoclonal antibody (1:10,000, Dia-An Biotechnology, catalog no. 2064); Mouse anti-HA monoclonal antibody (1:5000, Dia-An Biotechnology, catalog no. 2063); Mouse anti-β-actin monoclonal antibody (1:5000, Dia-An Biotechnology, catalog no. 2060); Mouse anti-GAPDH monoclonal antibody (1:1000, Santa Cruz, sc-47724); Histone H3 antibody (1:1000, Santa Cruz, sc-517576); Rabbit anti-MITA/STING polyclonal antibody (1:5000, Proteintech, catalog no. 19851-1-AP); Rabbit anti-IRF3 polyclonal antibody (1:1000, Proteintech, catalog no. 11312-1-AP); Rabbit anti-TBK1 monoclonal antibody (1:1000, Cell Signaling Technology, catalog no. 3504); Rabbit anti-phospho-IRF3 (S386) monoclonal antibody (1:1000, Abcam, AB76493); Rabbit anti-phospho-TBK1 (S172) monoclonal antibody (1:1000, Cell Signaling Technology, catalog no. 5483); Rabbit anti-IRF1 monoclonal antibody (1:1000, Cell Signaling Technology, catalog no. 8478); Rabbit IgG (Proteintech, catalog no. 20010049); Mouse anti-ICP0 monoclonal antibody (1:1000, Santa Cruz, sc-53070); Mouse anti-ICP8 monoclonal antibody (1:1000, Santa Cruz, sc-53329); Mouse anti-ICP27 monoclonal antibody (1:1000, Santa Cruz, sc-69806); Mouse anti-ICP5 monoclonal antibody (1:1000, Santa Cruz, sc-56989); IRDye 800CW Goat anti-Rabbit and Goat anti-Mouse secondary antibodies (1:10,000, LI-COR); Anti-FLAG beads (Dia-An Biotechnology); Protein A/G agarose (GE healthcare).

Techniques: Transduction, Control, Infection, Expressing, Quantitative RT-PCR, Labeling, Western Blot, Transfection, Plasmid Preparation, Luciferase, Stable Transfection

Journal: Virologica Sinica

Article Title: Nuclear translocation of Sp100A suppresses multiple DNA viruses and limits HSV-1 lytic replication in vivo

doi: 10.1016/j.virs.2026.02.003

Figure Lengend Snippet: Modulation of viral replication by Sp100A mutations on HSV-1. A HEp-2 cells were mock infected, or with the indicated viruses at an MOI of 5 for 6 h. Protein expression was evaluated by immunoblotting with anti-Flag, anti-β-actin, anti-TK, anti-ICP0, and anti-ICP8 antibodies. Note that the target proteins (Flag, TK, ICP0, and ICP8) were detected on separate blots. β-actin was used as a loading control and was run on a parallel blot using the same batch of protein lysates and identical loading amounts to confirm equal sample loading. B, C Growth kinetics of the indicated viruses in HEp-2 cells at MOIs of 0.01 ( B ) and 5 ( C ). D, E Growth kinetics of the indicated viruses in Vero cells at MOIs of 0.01 ( D ) and 5 ( E ). Data in B – E , statistical comparisons of viral titers were performed between group rHSV-1 and groups rHSV-1-A, rHSV-1-A S188A and rHSV-1-A S188D , as well as between group rHSV-1-A S188A and groups rHSV-1-A and rHSV-1-A S188D . Statistical analyses were performed using two-way ANOVA. F Plaque size comparison of the indicated viruses in HEp-2 and Vero cells. G, H Plaque size of each virus was quantified from 50 plaques using ImageJ and plotted. The data were normalized to the mean plaque size of the rHSV-1-A S188A group. Statistical analyses were performed using one-way ANOVA. Data presented as mean ± SD. ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Antibodies and drugs used in this study included the following: rabbit polyclonal anti-Flag antibody (Catalog No. 80010, Proteintech, USA), rabbit polyclonal anti-Sp100 antibody (Catalog No. GTX131569, GeneTex, USA), mouse monoclonal anti-Flag antibody (Catalog No. AB0008; Abways, Beijing, China), mouse monoclonal anti-β-actin antibody (Catalog No. 1000166; Sino Biological, Beijing, China), rabbit monoclonal anti-GAPDH antibody (Catalog No. AB0037, Abways Technology, Beijing, China), mouse monoclonal anti-Histone H3 antibody (Catalog No. 100005-MM01, Sino Biological, Beijing, China), rabbit polyclonal anti-TSG101 antibody (Catalog No. 28283-1-AP; Proteintech; USA), mouse monoclonal anti-calnexin antibody (Catalog No. sc23954, Santa Cruz, USA), mouse monoclonal anti-ICP0 antibody (Catalog No. sc53070, Santa Cruz, USA), mouse monoclonal anti-ICP8 antibody (Catalog No. ab20194, Abcam, UK), rabbit polyclonal anti-TK antibody (custom-made, GenicBio Limited, Shanghai, China), Alexa Fluor 594-labeled goat anti-rabbit IgG (H + L) (Catalog No. 2165334, Invitrogen, USA), goat anti-mouse IgG-HRP (Catalog No. 31430, Invitrogen, USA), goat anti-rabbit IgG (H + L)-HRP (Catalog No. 32460, Invitrogen, USA), and a protease inhibitor cocktail (Catalog No. EO0492, Thermo Scientific, USA).

Techniques: Infection, Expressing, Western Blot, Control, Comparison, Virus

Journal: bioRxiv

Article Title: Identification of Viral Activators of the HSV-2 UL13 Protein Kinase

doi: 10.1101/2025.07.06.663391

Figure Lengend Snippet: Line 1, wild-type HSV-2 186 genome; Line 2, domain of the UL12 gene to the UL15 gene; Line 3, domain of the UL13 gene; Lines 4 to 8, recombinant viruses with mutations in the UL13 gene; Line 9, domains of UL54 (ICP27) to UL56 and Us9 to Us11 genes; Line 10, domains of the UL55 and Us10 genes; Lines 11 to 16, recombinant viruses with mutations in the UL55 and/or Us10 genes.

Article Snippet: Antibodies and dilutions used in immunoblotting were as follows: commercial mouse monoclonal antibodies to Flag-tag (M2; Sigma, 1:1000), Strep-tag II (4F1; MBL, 1:1000), HA-tag (TANA2; MBL, 1:1000), β-actin (AC15; Sigma, 1:1000), ICP27 (H1142; Santa Cruz, 1:2000) and rabbit polyclonal antibodies to VP23 (CAC-CT-HSV-UL18; Cosmo Bio, 1:2000), UL37 (CAC-CT-HSV-UL37; CosmoBio, 1:2000) green fluorescent protein (GFP) (598; MBL; 1:1000).

Techniques: Recombinant

Journal: mBio

Article Title: Sp1 facilitates continued HSV-1 gene expression in the absence of key viral transactivators

doi: 10.1128/mbio.03479-23

Figure Lengend Snippet: Antibodies for immunoblot experiments

Article Snippet: ICP0 , East Coast Bio , H1A027.

Techniques: Western Blot