In vitro assessment of RAS GAP activity
To estimate RAS GAP activity, HA-H-RAS together with wild-type NF or
variant NF were expressed in COS-7 or HEK 293T cells. GTP-bound RAS was
subsequently isolated using glutathione-agarose beads coated with
recombinant GST-RAF-RBD [van Triest et al., 2001]. See Supplementary
Information for details.