Journal: The Journal of Biological Chemistry

Article Title: Protein tyrosine kinase Src suppresses hepatitis C virus particle release through regulation of Ndrg1

doi: 10.1016/j.jbc.2025.111125

Figure Lengend Snippet: Tyrosine kinase inhibitors differentially modulate intra- and extracellular HCV infectious particle production . A , schematic overview of the experimental workflow. Huh-7.5 cells were seeded 1 day prior to infection (d-1), infected with HCVcc (MOI = 5) and treated with TKIs on day 0 (d0). After 24 h, the medium was refreshed and TKIs were re-administered (d1). After an additional 48 h of incubation (d3), culture supernatants and cell lysates were collected for TCID 50 assays and RT-qPCR. B , Quantification of extracellular HCV infectivity following TKI treatment. Naïve Huh-7.5 cells were inoculated with supernatants collected 72 h post-infection from TKI-treated cultures. Infectious titers were determined by TCID 50 assays using immunostaining for the HCV core protein. C , Extracellular HCV RNA was extracted and quantified by RT-qPCR. Viral RNA copy numbers were normalized to copies per microliter of supernatant. D , Comparison of intracellular and extracellular HCV titers following treatment with selected TKIs. Relative changes are shown for Bosutinib ( left ), Entospletinib ( middle ), and BAY 61-3606 ( right ) compared to untreated controls. Data are presented as mean ± SD from at least three independent biological replicates. Statistical significance was analyzed using One-way ANOVA followed by the Dunnett’s multiple comparisons test ( B , C ) or Mann-Whitney U test ( D ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: In addition, two Syk kinase inhibitors, Entospletinib (Selleck, S7523) and BAY 61-3606 (Selleck, S7006), and the Egfr inhibitor Gefitinib (Funakoshi, CS-0124) were also included in the analysis.

Techniques: Infection, Incubation, Quantitative RT-PCR, Immunostaining, Comparison, MANN-WHITNEY

Journal: The Journal of Biological Chemistry

Article Title: Protein tyrosine kinase Src suppresses hepatitis C virus particle release through regulation of Ndrg1

doi: 10.1016/j.jbc.2025.111125

Figure Lengend Snippet: Effect of tyrosine kinase inhibitors on HCV infection efficiency and viral protein expression . A , Huh-7.5 cells were infected with HCVcc (MOI = 5) and treated with Bosutinib, Entospletinib, or BAY 61-3606 following the protocol described in A . At 72 h post-infection, cells were fixed and stained with an anti-core monoclonal antibody ( top panels ) and Hoechst to visualize nuclei ( bottom panels ). Scale bar, 100 μm. B , quantification of intracellular HCV RNA levels treated with the indicated inhibitors. Total RNA was extracted and analyzed by RT-qPCR. Data represent the relative expression normalized to untreated controls. C , immunoblot analysis of HCV NS3 and core protein expression. Detergent-soluble lysates were prepared from infected cells treated with the indicated inhibitors, separated by SDS-PAGE, and probed with anti-NS3, anti-core, and anti-actin antibodies. Densitometric quantification of NS3 and core protein levels relative to actin is shown in the right panels . Data are presented as mean ± SD from at least three independent biological replicates. Statistical significance was analyzed using Welch’s t test. ns, not significant. IB, immunoblot.

Article Snippet: In addition, two Syk kinase inhibitors, Entospletinib (Selleck, S7523) and BAY 61-3606 (Selleck, S7006), and the Egfr inhibitor Gefitinib (Funakoshi, CS-0124) were also included in the analysis.

Techniques: Infection, Expressing, Staining, Quantitative RT-PCR, Western Blot, SDS Page