Fig. 1: RSV-F protein accumulates in cholesterol-rich lysosomes in infected cells. | Nature Communications

Fig. 1: RSV-F protein accumulates in cholesterol-rich lysosomes in infected cells.

From: Cholesterol-rich lysosomes induced by respiratory syncytial virus promote viral replication by blocking autophagy flux

Fig. 1

HEp-2, 16HBE, or HBECs cells were either mock-infected or infected with RSV (MOI = 1) in the presence or absence of U18666A (10 μM) for the indicated durations. a–f Immunocolocalization of RSV proteins (RSV-F, RSV-G, and RSV-N), cholesterol (filipin III), and LAMP-1 in mock-infected, RSV-infected (24 h post-infection), and U18666A-treated cells (HEp-2 (a, b); 16HBE (c, d); HBECs (e, f). Scale bar: 10 μm. Data (n = 20 micrographs) are one representative of three independent experiments. g Western blotting analysis of RSV-F protein in lysosomes. h The cholesterol content in HEp-2 cells 0 h, 1 h, 2 h, 6 h, 12 h, 24 h, and 48 h after RSV infection was determined using an Amplex™ Red Cholesterol Assay Kit (n = 3 independent experiments). Image parameters: Scaling-per Pixel (a, c, e: 0.032 × 0.032 μm2); Image size-pixels (a, c, e: 2432 × 2432); image size-scaled (a, c, e: 78.01 × 78.01 μm2); objective (a, c, e: plan-apochromat 63×/1.40 oil DIC M27); scan zoom (a, c, e: 1.3). Data are shown as the mean ± SD, statistical analysis using one-way ANOVA (####P < 0.0001 compared to the blank control group; ****P < 0.0001 compared to the viral control group).

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