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Fig. 3: Rspo2 is a negative feedback inhibitor in the BMP4 synexpression group. a In situ hybridization of rspo2 and bmp4 in Xenopus laevis at gastrula (St. 11, dorsal to the top, vegetal view), neurula (St. 15, anterior to the top, dorsal view), and tadpole (St. 32, anterior to the left, lateral view). Dashed lines, dorsal blastopore lip (dbl); anp, anterior neural plate; di, diencephalon; mhb, mid-hindbrain boundary; ov, otic vesicle; a, atria; nt, neural tube; de, dorsal eye; h, heart; pdm, proctodeum. Scale bar, 0.5 mm. b Microinjection and experimental scheme for c–e. 2 or 4 cell stage Xenopus laevis embryos were animally injected with control (ppl) or bmp4 mRNA. The animal cap (AC) explants were dissected from injected embryos at stage 8, and either treated or untreated with cycloheximide (CHX) until control embryos reached stage 10 for qRT-PCR (c) or in situ hybridization (d, e). c qRT-PCR of rspo2, sizzled, and vent1 expression in the AC explants injected and treated as indicated. Data are pooled from three independent experiments with similar results and displayed as means ± SD. ns, not significant, *P < 0.05, **P < 0.01, ****P < 0.0001 from two-tailed unpaired t-test. d In situ hybridization of rspo2 and sizzled in the AC explants injected and treated as indicated. e Quantification of (d). n = number of the AC explants. f Model for Rspo2 function as a negative feedback inhibitor of BMP4 in Xenopus dorsoventral patterning. |
