Extended Data Fig. 7: Cleavage of NEMO in cell extracts.
a, Cell lysates of the human brain endothelial cell line hCMEC/D3 were incubated with Mpro (25 µM) for different durations and blotted against NEMO and Mpro (representative for at least 3 experiments at different conditions). b, HEK293T cells were transfected with a plasmid encoding for the mouse NEMO linked to GFP via a 2A sequence. After cell lysis, Mpro (25 µM) was added and incubated for the indicated durations. Antibodies against the 2A linker were used to identify NEMO and its cleavage products (representative for 2 experiments). c, hCMEC/D3 cells were transfected with the same plasmid as described in b (pNEMO-2A-GFP) and additionally with an empty control plasmid or a plasmid containing the HA-tagged form of the SARS-CoV-2 Mpro protease. After 2 days, cells were lysed and blotted against NEMO, 2A, Mpro, and actin (representative for at least 9 experiments). The red-boxed part of the blots is shown in Fig. 3c.
