Supplementary Figure 1: Generation of SnapCaaX lines.
(a) Schematic diagram of the wild type Rosa26 locus. (b) The Ai strategy and its recombination product. Cre-mediated removal of the stop cassette induces expression. Also included are PhiC31 recognition sites AttB/AttP for removal of the PGK-Neo, and FRT sites to allow for Flp recombinase–mediated replacement as described for the original Ai9 vector. (c) The FLEx strategy and its recombination product. Cre-mediated recombination induces inversion and then excision of a stop cassette (d) Southern blot of positive and negative ES clones targeted using the Ai strategy. (e) Southern blot of positive and negative ES clones targeted with the FLEx strategy. (f) SNAPCaaX transcript levels in the Ai and FLEx targeting strategies. (g) In-gel fluorescence of protein extracts labelled with 505-Star. Lane 1, Control. Lane 2, FLEx. Lane 3, Ai. A western blot of actin as loading control is shown below.
