Fig. 2.

In vivo efficacy of caged Rockout. (A-C) Stage 39 Xenopus embryos were exposed to 40 μM cRO for 2 hours, rinsed and individually irradiated on the right-hand side of the prospective gut (A); green-to-red photoconversion of EosFP indicates the decaged region (B, ventral view; C, right view). (D-I) Irradiated groups were then cultured in embryo medium (0.1× MMR) in the dark until the end of gut morphogenesis (stage 46). Embryos grown in the dark in 0.1× MMR (untreated, D), DMSO (F) or cRO (H) have long coiled guts, compared with those cultured in 30 μM RO (E), which have uniformly straight, un-elongated guts. Right side UV irradiation does not affect gut morphology in DMSO controls (G), but induces regions of defective elongation on the right side of the gut (arrowheads) in cRO-exposed embryos (I). (J-O) The proportion of normal (J), mild (K) and severe (L) gut elongation defects induced by decaging of cRO is dependent on the concentration of cRO to which the embryos are exposed (120 minutes uptake, 60 seconds irradiation; M), the uptake time (15 μM cRO, 60 seconds irradiation; N), and the length of UV irradiation (15 μM cRO, 120 minutes uptake; O). Control embryos may be exposed to UV for up to 2 minutes or cRO for up to 240 minutes (non-irradiated) without adverse effect (not shown).