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. 2022 May 8;23(6):e54305. doi: 10.15252/embr.202154305

Figure 4. ADAM10 is required for lung cell syncytia formation.

Figure 4

  1. Experimental scheme of syncytia formation assay. Donor cells (HEK293T) co‐expressing the SARS‐CoV‐2 spike protein along with GFP were co‐cultured with A549‐ACE2 NTC or KO acceptor cells. After 24 h, co‐cultures were analyzed by confocal microscopy.
  2. Representative images of syncytia formation assay. Large syncytia were observed for NTC and ADAM17 KO cells but were strongly reduced when ADAM10 was knocked out, either alone or together with ADAM17 or upon inhibition with BB94 (10 µM). For both ADAMs, the cell lines obtained with gRNA sequence 1 were used. Images show GFP fluorescence and Hoechst staining. Scale bars, 50 µm.
  3. Quantification of images from fluorescence microscopy (B). Left plot: the area of the fused cells in green (GFP) was quantified and normalized to the Hoechst signal (blue) of the entire image to account for differences in cell density. Right plot: The nuclei within syncytia were determined by calculating the ratio between the Hoechst signal within syncytia and the total Hoechst signal in the entire image. The data are normalized to NTC DMSO (N ≥ 8). Two‐way ANOVA with Tukey’s correction for multiple comparisons. All data are represented as mean ± 95% CI of at least three independent experiments. **P < 0.01, ***P < 0.001. See also Fig EV4.

Source data are available online for this figure.