Journal: Materials Today Bio
Article Title: Advancing influenza virus treatment: in vitro and ex vivo studies of PI3K inhibitor-loaded lipid nanoparticles
doi: 10.1016/j.mtbio.2025.102587
Figure Lengend Snippet: In the presence of pictilisib and POx-Lipid (pictilisib) IAV replication is reduced in ex vivo mouse lung slices. (A) Mouse lung was extracted, cut into slices, infected with A/H1N1pdm09 Jena5258 for 3 h (B–D) or left uninfected (C–D) and afterwards treated with 0.5 μM pictilisib, POx-Lipid (pictilisib) or respective volumes of the controls (DMSO, POx-Lipid) for 45 h. (B) At 48 h p.i. supernatants were used to determine progeny virus titers (PFU mL −1 ) by plaque assay. The mean +SD of three independent mice with four technical replicates is depicted. Statistical significance was determined using one-way ANOVA with Dunnett's multiple comparisons test. ∗∗ p < 0.01. Comparisons without asterisks did not reach statistical significance. (C–D) Alveolar macrophages were stained with a rat anti-CD68 primary antibody and an anti-rat Cy3 secondary antibody (yellow). Viral nucleoprotein (NP) is stained with a mouse anti-IAV NP primary and an anti-mouse AF488 secondary antibody (cyan). Lung structure is shown through brightfield imaging. Scale bars represent 200 μm. Depicted is one out of three independent mice. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Article Snippet: At 48 h p.i., the slices were fixed for 1 h in 3.7 % formaldehyde (FA; Sigma-Aldrich, Darmstadt, Germany) at RT, permeabilized with 0.1 % Triton-X-100 for 1 h at RT and blocked with 10 % normal goat serum (NGS; Sigma-Aldrich, Darmstadt, Germany) for 1 h. Subsequently, the primary antibodies mouse anti-IAV nucleoprotein (NP; MCA400; BioRad, Berkeley, CA, USA), rabbit anti-IAV M2 (GTX125951; GeneTex, Downers Grove, IL, USA) and anti-rat CD68 (14-0681-82; Thermo Fisher Scientific, Schwerte, Germany) were added at a ratio of 1:200 for 1 h at RT.
Techniques: Ex Vivo, Infection, Virus, Plaque Assay, Staining, Imaging
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