Journal: iScience

Article Title: Virus-host interactome reveals host cellular pathways perturbed by tick-borne encephalitis virus infection

doi: 10.1016/j.isci.2026.115821

Figure Lengend Snippet: TBEV NS5 interacts with P300 to modulate G0/G1 cell cycle progression (A) Cells were transfected with the plasmids of Flag-tagged EV and ten TBEV viral proteins, the cell distribution was analyzed by flow cytometry. (B) The expression plasmids of Flag-EV, 0.5 and 1.0 μg Flag-NS5 were transfected into A549 cells, the cell distribution was analyzed by flow cytometry. The experiment was repeated for three times, and the percentages of cells were shown in column graph. (C) Cells were transfected with Flag-EV or Flag NS5 together with HA-P300, the interaction between NS5 and P300 was analyzed by co-immunoprecipitation analysis. (D) The colocalization of P300 (green) and NS5 (red) were examined by immunofluorescence analysis. Scale bars, 10 μm. (E–G) Cells transfected with NS5 were collected after 48 h, the expression of CDK4, CDK6, and P16 was analyzed by immunoblot (E). (F) Quantification of protein levels from (E). Data are normalized to actin and presented as fold change relative to control (mean ± SD, n = 3). (G) qPCR analysis of mRNA expression of CDK4, CDK6, and P16 upon NS5 overexpression. Data are normalized to GAPDH using the 2ˆ(-ΔΔCt) method and shown as fold change relative to control (mean ± SD, n = 3). (H) A549 cells transfected with HA-EV, HA P300 and P300 Hm were further transfected with NS5, the distribution of cells was analyzed by flow cytometry. The experiments were repeated for three times, and the proportions of cells were shown in column graph, the expression of indicated proteins was assessed by immunoblot assay. (I) Cells transfected with P300 siRNA and NC siRNA (siNC) were further transfected with NS5, the distribution of cells was analyzed by flow cytometry, and the proportions of cells and the relative expression of P300 mRNA were shown in column graph. (J) Cells transfected with HA-EV, HA P300, and P300 Hm were mock-infected or infected TBEV, the distribution of cells was analyzed by flow cytometry. (K) Cells transfected with P300 siRNA and NC siRNA were mock-infected or infected TBEV, the distribution of cells was analyzed by flow cytometry. (L) Outline of G0/G1 cell-cycle arrest regulated by TBEV NS5. Data in column graphs are represented as mean ± SEM of three independent experiments. The p values are calculated and reported using one-way ANOVA.

Article Snippet: For small interfering RNA (siRNA) targeting P300, 5′-GGACUACCCUAUCAAGUAATT-3′, was synthesized at Sangon Biotech (Shanghai, China).

Techniques: Transfection, Flow Cytometry, Expressing, Immunoprecipitation, Immunofluorescence, Western Blot, Control, Over Expression, Infection

Journal: iScience

Article Title: Virus-host interactome reveals host cellular pathways perturbed by tick-borne encephalitis virus infection

doi: 10.1016/j.isci.2026.115821

Figure Lengend Snippet: P300 inhibitors and CDK4 agonist restrict viral gene expression by interfering with TBEV-induced cell-cycle arrest (A and B) A549 cells were either mock-infected or infected with TBEV, the cells were then treated with indicated concentration of C646 (A) and CPI-637 (B), the supernatants were collected at 48 hpi, and the mRNA level of TBEV E gene was analyzed by probe qPCR. (C–F) Cells mock-infected or infected with TBEV at the MOI of ten were treated with 10 μM C646 and CPI-637, the expression of TBEV NS1 protein was detected by immunoblot (C and D) and immunofluorescence analysis (E), and the cell cycle distribution was analyzed by flow cytometry (F). Scale bars, 50 μm. (G) Cells transfected with HA-EV, HA P300, and P300Hm were infected TBEV, the mRNA level of TBEV E gene in the supernatants was analyzed by probe qPCR. (H) Cells transfected with P300 siRNA and NC siRNA were infected TBEV, the mRNA level of TBEV E gene in the supernatants was analyzed by probe qPCR. (I) A549 cells were treated with the indicated concentrations of chrysin for 2 h, the cells were then infected with TBEV, the supernatants were collected at 48 hpi, and the mRNA level of TBEV E gene was analyzed by probe qPCR. (J–L) Cells were treated with 10 μM chrysin for 2 h, the cells were then mock-infected or infected with TBEV at the MOI of ten, the expression of TBEV NS1 protein was detected by immunoblot (J), and the cell distribution was analyzed by flow cytometry (K). The percentage of cells (L) upon chrysin treatment was shown in column graphs. Scale bars, 50 μm. Data in column graphs are represented as mean ± SEM of three independent experiments. The p values are calculated and reported using one-way ANOVA.

Article Snippet: For small interfering RNA (siRNA) targeting P300, 5′-GGACUACCCUAUCAAGUAATT-3′, was synthesized at Sangon Biotech (Shanghai, China).

Techniques: Gene Expression, Infection, Concentration Assay, Expressing, Western Blot, Immunofluorescence, Flow Cytometry, Transfection

Journal: Advanced Science

Article Title: β‐Elemene Rescues Radiation‐Induced Enteritis by Orchestrating a Host‐Microbiome Circuit That Fuels Epigenetic DNA Repair

doi: 10.1002/advs.202521445

Figure Lengend Snippet: EP300 Functions as the Lactyl‐transferase for RBBP4 to Establish a Positive Feedback Loop (A) Molecular docking model. Left: Overall predicted structure of the interaction between a lactylated RBBP4 peptide (light blue) and the EP300 protein (cyan). Right: Magnified view showing that the K26 lactylation (Lys 26) on RBBP4 can form hydrogen bonds with key residues such as Glutamic Acid (GLU 1519) in the EP300 catalytic domain, thereby stabilizing the interaction. (B,C) Western blot (WB) analysis. (D,E) Co‐immunoprecipitation (Co‐IP) experiments with IP performed using an anti‐RBBP4 antibody (D) and an anti‐EP300 antibody (E). The results indicate that The EP300 agonist CTB enhanced (D), while the inhibitor C646 reduced (E), the interaction between EP300 and RBBP4. (F) Multiplex immunofluorescence staining of mouse ileal tissues. Compared to the radiation‐only group (IR+PBS), the β‐elemene treatment group (IR+β‐elemene) showed significantly enhanced expression and co‐localization of MCT1 (cyan), CD147 (green), RBBP4‐K26la (yellow), POLD1 (red), and POLD3 (purple) in the intestinal crypt regions. (G) Summary and schematic model of the mechanism. This study elucidates that β‐elemene promotes cellular uptake of L. gasseri ‐derived lactate by enhancing the membrane translocation of MCT1 in a CD147‐dependent manner. Intracellular lactate, through the lactylation of RBBP4 at the K26 site, recruits EP300 to the promoter regions of downstream genes ( POLD1 / POLD3 ), catalyzing H3K27ac modification. This ultimately activates the DNA damage repair pathway, protecting intestinal epithelial cells from radiation injury. Original full and unaltered blots can be found in Figure .

Article Snippet: The EP300 inhibitor C646 and agonist CTB were purchased from MedChemExpress.

Techniques: Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Multiplex Assay, Immunofluorescence, Staining, Expressing, Derivative Assay, Membrane, Translocation Assay, Modification