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. 2020 Jul 22;9:e56907. doi: 10.7554/eLife.56907

Figure 2. IAV-induced barrier damage in high glucose condition is dependent upon the presence of endothelial cells.

(A) Schematic representation of the in vitro mono-culture model of the alveolar epithelial-endothelial barrier. Image created with Biorender. (B) Left: Measurement of epithelial mono-culture barrier integrity using TEER (Ω) readings following infection with medium (‘mock’) or IAV. Data are expressed relative to the baseline TEER and the TEER of mock-infected cells at each time point. Statistical significance was determined using a two-way ANOVA with a Bonferroni post-test. Right: Permeability of epithelial mono-culture to FITC-dextran 24 hr post-infection. Data show the percentage of FITC detected in the lower compartment relative to mock-infected wells (defined as 0). Infected wells that had less detected FITC than mock-infected were set to 0. Statistical significance was determined using a Student’s unpaired t-test. (C) Left: PFU/mL of IAV detected in the lower compartment and upper compartment 24 hr post-IAV infection. A dashed line indicates the detection limit of the assay. Right: mRNA detected by qPCR in epithelial cells 24 hr post-IAV infection. Viral replication represented as viral copy. Statistical significance was determined using a Student’s unpaired t-test. (D) Percentage release of LDH from mono-culture epithelial (upper compartment) cells at 24 hr post-infection. Statistical significance was determined using a Student’s t-test. All data are pooled from a minimum of three independent experiments (with six biological replicates per group) and are shown as mean ± SEM. *: p<0.05.

Figure 2.

Figure 2—figure supplement 1. Endothelial cells provide a stabilising effect on the integrity of the epithelial barrier.

Figure 2—figure supplement 1.

Baseline TEER (Ω) of epithelial-endothelial co-cultures and epithelial mono-cultures following 3–4 days of media changes (prior to IAV infection). Data are pooled from a minimum of four independent experiments (each with 24 biological replicates per group) and shown as mean ± SEM. Statistical significance was determined using a Mann-Whitney test. *: p<0.05.