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. 2015 May 26;120:101–111. doi: 10.1016/j.antiviral.2015.05.008

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Copyright © 2015 Elsevier B.V. All rights reserved.

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Fig. 2 — Mouse lung slices support influenza virus replication. (A) To determine the virus production of lung slices of different thickness, 125-, 250-, 500- and 1000-μm thick lung slices were exposed to 200 μl of 10⁵ PFU/ml of PR8 (H1N1) for 48 h, and the supernatants were used to determine the virus titre by a TCID₅₀ assay. Comparisons between groups were performed by an unpaired t-test. Data are expressed as the means ± SEM. n.s., non-significant. *, 0.01 < p < 0.05 in comparison to 125-μm thick slices. (B–E) To determine the virus yields at different time points post infection, 250-μm thick slices were infected with 200 μl of 10⁵ PFU/ml PR8 (H1N1) (B, D) or HUBEI (H3N2) (C, E) virus. The supernatants were collected at indicated time points to perform the NA activity assay (B and C) and TCID₅₀ assay (D and E). (F) Infection of lung slices was assessed by the IFA assay. Slices with a thickness of 150 μm were exposed to 200 μl of 10⁶ PFU/ml PR8 (H1N1) or HUBEI (H3N2) viruses for 24 h. The slices were fixed with 4% paraformaldehyde and analysed by IFA. The images were captured using a Nikon multiphoton Confocal Microscope A1 MP⁺. The virus diluent was used as a negative control (NC) in the infection. Scale bar = 1000 μm. The right panels show enlarged regions of interest from the left panels.

Fig. 2 — Mouse lung slices support influenza virus replication. (A) To determine the virus production of lung slices of different thickness, 125-, 250-, 500- and 1000-μm thick lung slices were exposed to 200 μl of 10⁵ PFU/ml of PR8 (H1N1) for 48 h, and the supernatants were used to determine the virus titre by a TCID₅₀ assay. Comparisons between groups were performed by an unpaired t-test. Data are expressed as the means ± SEM. n.s., non-significant. *, 0.01 < p < 0.05 in comparison to 125-μm thick slices. (B–E) To determine the virus yields at different time points post infection, 250-μm thick slices were infected with 200 μl of 10⁵ PFU/ml PR8 (H1N1) (B, D) or HUBEI (H3N2) (C, E) virus. The supernatants were collected at indicated time points to perform the NA activity assay (B and C) and TCID₅₀ assay (D and E). (F) Infection of lung slices was assessed by the IFA assay. Slices with a thickness of 150 μm were exposed to 200 μl of 10⁶ PFU/ml PR8 (H1N1) or HUBEI (H3N2) viruses for 24 h. The slices were fixed with 4% paraformaldehyde and analysed by IFA. The images were captured using a Nikon multiphoton Confocal Microscope A1 MP⁺. The virus diluent was used as a negative control (NC) in the infection. Scale bar = 1000 μm. The right panels show enlarged regions of interest from the left panels.