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. 2024 Nov 22;14:28961. doi: 10.1038/s41598-024-79397-w

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — LCMV vRNA is present alongside NP in TNT-like connections. (A) At 24 hpi rLCMV infected A549 cells at an MOI of 1 were fixed and then stained for NP (red). Subsequently, coverslips were hybridised overnight with 48 specific FISH probes designed to hybridize with LCMV S vRNA (green). Widefield microscopy was then utilised to visualise NP and S vRNA within TNT-cell connections. Cell nuclei were DAPI stained. Uninfected cells and infected cells are shown. A zoomed-in region of interest (white box in the central row) is shown on the bottom row. (B) Line scan analysis of the region of interest in the zoomed merged image (dashed line in the bottom right panel of (A)) revealed peaks in intensity of NP and S vRNA were similar. (C) Co-occurrence analysis using the Manders coefficient method was performed for 10 TNT-like connections. The percentage of LCMV NP to S vRNA (Red) and S vRNA to LCMV NP (green) is shown, and individual images plotted.