ec 50 values calculation  (GraphPad Software Inc)

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Network pharmacology and targets analysis of baicalein against PRRSV . A Interaction analysis of target genes between baicalein and PRRSV-related. B PPI network analysis. 90 potential targets were input into a String database, and a PPI network was constructed ( P -value < 1.0 e−16 ). C Topological analysis by Cytoscape 3.10.0. The seven core targets were screened and highlighted in the right box. The core targets screening criteria were set as degree > 31. D GO enrichment analysis of the 90 potential targets. E Top 10 of KEGG signalling pathway analysis of the 90 potential targets. The size of the dot represents the counts of proteins in this pathway; the colour of the dot corresponds to different P -value ranges.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Construct

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Binding of baicalein with potential targets

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Binding Assay

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Molecular docking and MD stimulation of baicalein with EGFR . A Active pocket of EGFR prediction. The volume of the EGFR binding pocket was calculated with CHECOM algorithm by Yinfo technology. Active pocket of EGFR was shown in red with 25,465 Å 3 . B Molecular docking of EGFR (Uniprot ID: A0A5G2QBP1) with baicalein (Compound CID: 5281605) in a 3D model and 2D ligand interaction diagram. C Root Mean Square Deviation (RMSD) of baicalein and EGFR during molecular dynamics simulation. D Hydrogen bond numbers between baicalein and EGFR at different times. E Molecular docking of baicalein with another six core protein targets. E1-E6: SRC, TP53, MMP9, STAT3, INS, and ESR1.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Binding Assay

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Antiviral effect of baicalein on PRRSV infection in vitro . A 3D chemical structure of baicalein. B 50% cytotoxic concentration (CC 50 ) analysis. C Copies number of 10 –1 , 10 –2 , 10 – 3 fold dilution PRRSV. D CPEs of MARC-145 cells. MARC-145 cells were treated with DMSO, baicalein (80 μg/mL), and ribavirin (120 μM), respectively, and infected with PRRSV (100 TCID 50 ). PBS was used as a mock infection in the negative group. Scale bar = 100 μm. E The antiviral effect of baicalein on PRRSV was examined through the calculation of viral copies at various concentration gradients. F Immunofluorescence assay of protein N. Scale bar = 100 μm. G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0. The data are expressed as mean ± SD. Each experiment was performed with three biological replicates and three technical replicates. H Mean fluorescence intensity (Mean ± SEM, n = 3). * P < 0.05; ** P < 0.01, *** P < 0.005, **** P < 0.0001 compared with the DMSO treatment group.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Infection, In Vitro, Concentration Assay, Immunofluorescence, Construct, Fluorescence

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein inhibited PRRSV proliferation in pre-, co-, and post-treatment modes in MARC-145 cells . A Schematic diagram of time-of-addition assay. Briefly, MARC-145 cells were infected with PRRSV (100 TCID 50 ), and the cells were treated with 80 μg/mL baicalein for different times before (pre-treatment), during (co-treatment) or post-infection (post-treatment), as shown in the schedules. At 48 hpi, the cells and supernatants were harvested for analysis. B PRRSV copy number calculation using absolute quantification. C PRRSV-ORF7 relative expression analysis using qRT-PCR. D Virus titres of PRRSV. TCID 50 of PRRSV was calculated by the Spearman-Kärber method. Each experiment was performed with three biological replicates and three technical replicates. The data are expressed as mean ± SD. *, P < 0.05; **, P < 0.01, ***, P < 0.005, ****, P < 0.0001 compared with the DMSO treatment group.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Infection, Quantitative Proteomics, Expressing, Quantitative RT-PCR, Virus

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein directly interacted with PRRSV . A Schematic diagram of experimental design. Briefly, PRRSV (100 TCID 50 ) was subjected to incubation with baicalein at different concentrations (20, 40, and 80 μg/mL) in essential medium for a duration of 1 h at 37 °C. Following this incubation, PRRSV was isolated from baicalein through the process of ultrafiltration. The retrieved PRRSV was then challenged to MARC-145 cells. Subsequently, at 48 hpi, the cells were collected for analysis. B PRRSV copy number calculation using absolute quantification. C PRRSV-ORF7 relative expression analysis using qRT-PCR. D Virus titres of PRRSV. TCID 50 of PRRSV were calculated by the Spearman-Kärber method. Each experiment was performed with three biological replicates and three technical replicates. The data are expressed as mean ± SD. * P < 0.05; ** P < 0.01, *** P < 0.005, **** P < 0.0001 compared with the only PRRSV treatment group. E Active pocket of Gp5/M prediction. The volume of the Gp5/M binding pocket was calculated with CHECOM algorithm by Yinfo technology. Active pocket of Gp5/M was shown in green. F Molecular docking of Gp5/M (PRRSV-2 VR 2332, model 1, alphafold2) with baicalein (Compound CID: 5281605) in a 3D model. G Root Mean Square Deviation (RMSD) of baicalein and Gp5/M during MD simulation. H Hydrogen bond numbers between baicalein and Gp5/M in 100 ns.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Incubation, Isolation, Quantitative Proteomics, Expressing, Quantitative RT-PCR, Virus, Binding Assay

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein improved growth performance and immunity of PRRSV-infected weaned piglets . A Schematic diagram of experimental design of baicalein inhibiting PRRSV infection in piglets. B Average daily gain (ADG) of PRRSV-infected piglets with baicalein treatment. C Antibody level of PRRSV-infected piglets with baicalein treatment. D - E Macroscopic lung lesion changes in PRRSV-infected piglets with baicalein treatment. Macroscopic lungs of piglets in each group were scored by a scoring system. F The lungs of piglets were collected, and sections were prepared to stain with haematoxylin and eosin (H&E). Black arrows indicate the histopathological alterations, including progressive loss of lung architecture, thickening of the interlobular septa, and infiltration of lymphocytes (Mean ± SEM, n = 3, scale bar = 100 μm).

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Infection, Staining

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein improved antiviral activity of PRRSV-infected piglets . A - B IFA analysis of protein N using a rabbit PRRSV N protein polyclonal antibody (1:100, Bioss, Beijing, China). Expression of the protein N is indicated with red arrows. Scale bar = 100 μm; local enlarged images are shown in the right, scale bar = 20 μm (Mean ± SEM, n = 3). C - D Expression of the PRRSV N protein in lung tissues was detected using western blotting (Mean ± SEM, n = 3). E Viral loads of PRRSV calculated by absolute quantitation. F - G Expression of PRRSV protein N in lung tissues was detected by IHC with a rabbit PRRSV N protein polyclonal antibody (1:100, Bioss, Beijing, China). Expression of the N protein is indicated with black arrows. Scale bar = 100 μm (Mean ± SEM, n = 3). * P < 0.05; ** P < 0.01 compared with the PRRSV group.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Activity Assay, Infection, Expressing, Western Blot, Quantitation Assay

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein improved anti-inflammatory and antioxidant activity of PRRSV-infected weaned piglets . A The relative expression of inflammatory cytokines in the lung. B The antioxidant capacity in serum, liver and lung. The data are expressed as mean ± SD ( n = 5). T-AOC Total antioxidant capacity, GSH-PX Glutathione peroxidase, MDA Malondialdehyde, CAT catalase, SOD Superoxide dismutase.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Antioxidant Activity Assay, Infection, Expressing

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Baicalein blocked PRRSV entry via the EGFR-PI3K-AKT pathway . A PRRSV copy number calculation using absolute quantification. B PRRSV-ORF7 relative expression analysis using qRT-PCR. C Virus titres of PRRSV. TCID 50 of PRRSV were calculated by the Spearman-Kärber method. Each experiment was performed with three biological replicates and three technical replicates. The data are expressed as mean ± SD. D Protein expression in the lung of PRRSV-infected piglets by western blotting. E Band intensities were subjected to semi-quantitative analysis employing ImageJ software, and statistical significance of the results was analysed utilizing GraphPad Prism software. β-actin served as loading control. F Phosphorylation EGFR expression in the lung of PRRSV-infected piglets by IHC. Strongly positive signals are presented with red arrows in PRRSV group and slightly positive signals are presented with black arrows in PRRSV + Bai group. Scale bar = 50 μm. G Area of p-EGFR expression (Mean ± SEM, n = 3). *, P < 0.05; **, P < 0.01, ***, P < 0.005, ****, P < 0.0001 compared with the only PRRSV treatment group.

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Quantitative Proteomics, Expressing, Quantitative RT-PCR, Virus, Infection, Western Blot, Software, Control, Phospho-proteomics

Journal: Veterinary Research

Article Title: Baicalein inhibits PRRSV through direct binding, targeting EGFR, and enhancing immune response

doi: 10.1186/s13567-024-01440-5

Figure Lengend Snippet: Scheme summarizing the inhibitory effect of baicalein on PRRSV infection . The underlying antiviral mechanisms of baicalein including binding to PRRSV structural protein to directly kill the virus, inhibiting EGFR-PI3K-AKT pathway to block PRRSV entry, and boosting host immunity to inhibit PRRSV infection. The figure was generated by BioRender with authorised consent for publication

Article Snippet: G EC 50 of baicalein against PRRSV was constructed by GraphPad prism 8.0.

Techniques: Infection, Binding Assay, Virus, Blocking Assay, Generated