Journal: Journal of Virology

Article Title: p53-mediated regulation of electron transport chain and nucleotide synthesis during Newcastle disease virus infection

doi: 10.1128/jvi.01576-25

Figure Lengend Snippet: p53-mediated ETC stabilization promotes NDV replication ( A and B ) A549 cells were transfected with siRNA to knock down TP53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 6 hpi. OCR was measured using the Agilent Seahorse XFe96 extracellular flux analyzer ( A ). Basal/maximal respiration and ATP production were analyzed using GraphPad software ( B ). ( C and D ) A549 cells were transfected with siRNA to knock down TP53 and negative control siRNA (siNC). Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 18 hpi. Intracellular ROS levels were quantified via flow cytometry using DCFH-DA staining ( C ), with fluorescence intensity analyzed by FlowJo software ( D ). ( E and F ) H1299 cells were transfected with HA-p53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 18 hpi. Intracellular ROS levels were quantified via flow cytometry using DCFH-DA staining ( E ), with fluorescence intensity analyzed by FlowJo software ( F ). ( G–I ) A549 cells were transfected with siRNA to knock down TP53 and negative control siRNA (siNC). Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 12 hpi and were treated with varying concentrations of ETC inhibitors (rotenone, antimycin A, and FCCP) during this period. NC represents cells treated with DMSO. Western blotting was employed to detect the protein levels of NDV-NP, TP53, and β-actin ( G ). Cell supernatant was collected and the viral titer was determined by plaque assay ( H and I ). ( J–L ) H1299 cells were transfected with HA-p53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 12 hpi and were treated with varying concentrations of ETC inhibitors (rotenone, antimycin A, and FCCP) during this period. NC represents cells treated with DMSO. Western blotting was employed to detect the protein levels of NDV-NP, HA, and β-actin ( J ). Cell supernatant was collected and the viral titer was determined by plaque assay ( K and L ). Data are presented as means from three independent experiments. ns, no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mouse monoclonal anti-β-actin (66009-1-Ig) and rabbit polyclonal anti-p53 (10442-1-AP) were purchased from Proteintech.

Techniques: Transfection, Knockdown, Infection, Software, Negative Control, Flow Cytometry, Staining, Fluorescence, Western Blot, Plaque Assay

Journal: Journal of Virology

Article Title: p53-mediated regulation of electron transport chain and nucleotide synthesis during Newcastle disease virus infection

doi: 10.1128/jvi.01576-25

Figure Lengend Snippet: Model of NDV-induced p53 stabilization supporting ETC activity and nucleotide synthesis for viral replication. In the presence of p53, cells adapt to virus-induced metabolic stress and maintain nucleotide pools, supporting both cell survival and viral replication. Without p53, cells become reliant on intact ETC; ETC disruption quickly depletes nucleotides and impairs viral replication. Thus, while p53 preserves cellular stability, viruses may exploit this environment to enhance their own replication.

Article Snippet: Mouse monoclonal anti-β-actin (66009-1-Ig) and rabbit polyclonal anti-p53 (10442-1-AP) were purchased from Proteintech.

Techniques: Activity Assay, Virus, Disruption