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Fig. 1. Expression of lin28a and lin28b in the X. tropicalis embryo. (A,B) Temporal expression profiles of lin28a and lin28b mRNA (A) and protein (B) in early cleavage to late gastrula stage embryos as assessed by Q-PCR and western blot, respectively. (C-P) Spatial expression of lin28a (C-H) and lin28b (I-P) as assessed by in situ hybridisation. (C,E,M) Vegetal view; (D,F,H,K,N) transverse sections, lateral view; (G,L,O) higher magnification of F, K and N, respectively; (J) vegetal view of I. Stages are indicated: stage 8, mid-blastula; stage 10 to 10.5, early gastrula; stage 12.5 to 13, late gastrula. dbl, dorsal blastopore lip; vbl, ventral blastopore lip; es, ectodermal superficial layer; bs, basal sensorial layer. (Q-V) Fluorescence microscopy of cryosections from the circumblastoporal region of early gastrula stage 10.5 embryos. (Q) Immunolocalisation of lin28a expression. Red arrows indicate nuclear enrichment. (R) Nuclear DAPI staining in same section as Q. (S) Merge of Q and R. (T) Immunolocalisation of lin28b expression. (U) Nuclear DAPI staining in same section as T. (V) Merge of T and U. (W) Expression levels of lin28a, lin28b and the ubiquitously expressed housekeeping gene ornithine decarboxylase (ODC) in dnFGR4-injected embryos as compared with control embryos by Q-PCR. Solid line represents mRNA levels in corresponding control embryos (set at 1). **P<0.005 (Student t-test), for expression in experimental embryos compared with control embryos. Error bars indicate s.d. Scale bars: 50 μm. |
