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. 2021 Jan 19;296:100306. doi: 10.1016/j.jbc.2021.100306

Figure 7.

Figure 7

SARS-CoV-2 spike protein endocytosis is reduced by CHC knockdown in VERO cells. A, lysates were made from VERO cells transfected with control siRNA or with an siRNA established to selectively knockdown the expression of CHC. Cell lysates were prepared and immunoblotted with the indicated antibodies. B, VERO cells, treated as in (A), were incubated with purified spike protein for 30 min at 4 °C. The cells were then transferred to 37 °C for the indicated time periods before being returned to ice. After acid wash, cells were fixed and stained with DAPI to reveal nuclei and with antibody selectively recognizing spike protein. Scale bars = 40 μm for the low-magnification images and 10 μm for the higher magnification insets on the right. C, the graph showing quantification of experiments performed as in (B). n = 15 from 3 independent experiments, mean ± SEM; unpaired t test; ∗∗∗∗p < 0.0001, n.s, not significant. ACE2, angiotensin-converting enzyme 2; CHC, clathrin heavy chain; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.