Journal: Frontiers in Immunology

Article Title: Baicalin Induces a Potent Innate Immune Response to Inhibit Respiratory Syncytial Virus Replication via Regulating Viral Non-Structural 1 and Matrix RNA

doi: 10.3389/fimmu.2022.907047

Figure Lengend Snippet: Oligonucleotides used for RT-qPCR.

Article Snippet: Anti-NS1, anti-NS2, and anti-M antibodies were produced by AtaGenix (Wuhan, China).

Techniques:

Journal: Frontiers in Immunology

Article Title: Baicalin Induces a Potent Innate Immune Response to Inhibit Respiratory Syncytial Virus Replication via Regulating Viral Non-Structural 1 and Matrix RNA

doi: 10.3389/fimmu.2022.907047

Figure Lengend Snippet: Baicalin demonstrates potent anti-RSV activity in vitro . (A) The cytotoxic influence of baicalin was determined by assessing cell viability by CCK-8 assay. HEp-2 cells were treated in triplicate with baicalin increased concentrations for 72 h. Absorbance at 570 nm in relation to that of mock treated control was plotted for evaluation of the TC 50 value. (B) Expression of viral transcripts of the NS2 gene was analyzed by RT-qPCR in HEp-2 cells infected with RSV (MOI = 3) and treated with the indicated doses of baicalin. Data were normalized to Actb expression. Results are shown as mean ± SD (n = 3); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 – ANOVA with Tukey’s post-hoc test. (C, D) Baicalin IC 50 was determined through plaque reduction assessment in HEp-2 cells infected with RSV in the presence of the indicated doses of baicalin (C) . The number of plaques was counted and plotted as the percentage of plaque inhibition in relation to untreated cells, after 72 h (D) .

Article Snippet: Anti-NS1, anti-NS2, and anti-M antibodies were produced by AtaGenix (Wuhan, China).

Techniques: Activity Assay, In Vitro, CCK-8 Assay, Control, Expressing, Quantitative RT-PCR, Infection, Inhibition

Journal: Frontiers in Immunology

Article Title: Baicalin Induces a Potent Innate Immune Response to Inhibit Respiratory Syncytial Virus Replication via Regulating Viral Non-Structural 1 and Matrix RNA

doi: 10.3389/fimmu.2022.907047

Figure Lengend Snippet: Baicalin affects transcriptional and translational regulation of RSV gene products. (A) Baicalin causes the downregulation of viral genes NS1 and NS2, but not others. Expression of indicated viral transcripts in lung homogenates of RSV-infected mice ± baicalin was quantified by RT-qPCR. Outcomes were normalized to Actb . (B) Immunoblot analysis of NS1, NS2, and M protein in lung homogenates of RSV-infected mice ± baicalin showed downregulation in the expression of these proteins after treatment with baicalin. Representative blots from 3 animals are shown. β-actin was immunobloted as a loading control. (C) Polysome occupancy of NS1, NS2, and M RNAs. Baicalin caused translational downregulation of viral M RNA. The decrease in polysome occupancy observed for NS1 and NS2 is attributable to the decrease in their total RNA. Data shown in (A , C) represents mean ± SD (n = 5); *** P < 0.001, **** P < 0.0001, ns , not significant – ANOVA with Tukey’s post-hoc test.

Article Snippet: Anti-NS1, anti-NS2, and anti-M antibodies were produced by AtaGenix (Wuhan, China).

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: Baicalin Induces a Potent Innate Immune Response to Inhibit Respiratory Syncytial Virus Replication via Regulating Viral Non-Structural 1 and Matrix RNA

doi: 10.3389/fimmu.2022.907047

Figure Lengend Snippet: Model summarizing mechanism underlying potent innate immune response elicited against RSV infection by treatment with baicalin. Translational regulation of RSV viral M RNA was previously elucidated . Briefly, baicalin suppresses translation of the viral M protein and transcription of the viral NS1 and NS2 proteins and downregulates the expression of pro-inflammatory cytokines while inducing expression of type I IFNs, thus mounting a potent anti-RSV innate immune response.

Article Snippet: Anti-NS1, anti-NS2, and anti-M antibodies were produced by AtaGenix (Wuhan, China).

Techniques: Infection, Expressing