Fig. 2
KRASG12V induction by incubation in cyclofen or photo-activation of caged cyclofen. (A,E) Injecting UAS:kRASG12V-T2A-CFP; ubi:Eos plasmid into Tg(ubi:Gal4-ERT2) embryos resulted in a mosaic expression of EosFP protein, but (in absence of free cyclofen) no expression of CFP. (B,F) Embryos treated with 2??M cyclofen at 32hpf display expression of CFP co-localized with EosFP expression (denoted by arrows). (C,G) Incubation of caged cyclofen (with no UV) did not activate CFP expression while (D,H) 2?min UV uncaging effectively activated the expression of CFP 12?hours later (denoted by arrows). The induction of kRASG12V by cyclofen was confirmed by kRASG12V mRNA in situ hybridization. Human kRASG12V mRNA was transcribed only in the embryo incubated in cyclofen (J), but not in the control embryo (I). Scale bar: 200 ?m.