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Fig. 2. lmx1b MO1 and MO2 prevent full size glomus formation. (A) lmx1b MO1 (panels b, e) or MO2 (panels h and k) was injected into the V2 blastomere at the 8-cell stage. All injections were carried out using a lineage tracer, either GFP (panels a–f) or LacZ (panels g–l). The size of the glomus was assessed by wt1 in situ hybridisation at stage 35/36. Injection of cMO does not induce any phenotype on the injected side (compare panels a to d and g to j). lmx1b MO1 injected embryos (5 ng) showed a reduced glomus (compare panel b to e). A similar phenotype was observed following the injection of lmx1b MO2 (10 ng) (compare panels h to k). Injection of lmx1b-mut mRNA (2.5 ng) can partially rescue the phenotype of MO1 (panels c and f) and MO2 (panels i and l). The injected side is marked with an asterisk. (B) lmx1b MO1 (panels b, e) or MO2 (panels h and k) was injected into one cell of 2-cell embryos. All injections were done using a lineage tracer, either GFP (panels a–f) or LacZ (panels g–l). The size of the glomus was assessed by nephrin in situ hybridisation at stage 33/34. Injection of cMO does not induce any phenotype on the injected side (compare panels a to d and g to j). lmx1b MO1 injected embryos (5 ng) showed a reduced nephrin-staining area on the injected side (compare panels b to e). A similar phenotype was observed following the injection of lmx1b MO2 (10 ng) (compare panels h to k). Injection of lmx1b-mut mRNA (2.5 ng) can partially rescue the phenotype of MO1 (panels c and f) and MO2 (panels i and l). The injected side is marked with an asterisk. |
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Fig. 6. Over-expression of lmx1b and its potential binding partners affects nephrin expression. mRNAs of lmx1b and its potential binding partners were injected either alone (panel A) or in combination (panel B), together with lacZ, into one cell of 2-cell embryos. Red Gal staining followed by nephrin in situ hybridisation was carried out at stage 33/34 in order to assess any glomus phenotype. For each embryo, the injected side (indicated by an asterisk, panels a–d) was compared to the uninjected side (panels e–h). Injection of lmx1b mRNA alone did not induce any significant change in nephrin staining (A, compare panel a to e), whereas injection of lim1 or ldb1 alone resulted in the formation of an enlarged and reduced nephrin domain respectively (A, b and f; A, c and g). These phenotypes can be partially rescued by the co-injection of lmx1b (B, b and f; B, c and g). Co-injection of lim1 and ldb1 rescued both phenotypes, the embryos displayed on the injected side a nephrin domain, similar in area to the uninjected side (B, a and e). Injection of E47 alone or in combination with lmx1b did not induce any statistically significant phenotype (A, d and h and B, d and h). |