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Figure 5. znf367 morphants analysis for proliferating and differentiating precursors (A) mRNA distribution of pcna in znf367 morphant and control. (B) Statistical analysis of the data in A. (C) RT-PCR analysis revealed a significant increase of pcna. (D,E) Znf367 depletion leads to a significant reduction of proliferating cells compared to the un-injected side. pH3 positive cells were counted in the areas defined by the black rectangles. Statistical evaluation of the data is shown (E). (F) RT-PCR analysis of cyclinB1 (ccnb1). The levels of expression for the indicated mRNAs were evaluated for Co-MO or ZNF367-MO embryos by qRT-PCR in triplicate, and normalized to glyceraldehyde 3-phosphate dehydrogenase (gapdh) expression. (G–G’) p27 is down regulated at stage 18 in znf367 morphants (arrowheads). (G) dorsal view; (G’) frontal view. (H–H’). p27 expression in control embryo, (H) dorsal view; (H’) frontal view. (I–L) rx1 and elrD expression at tailbud stages confirmed the phenotype observed at neurula stages: increased expression of rx1 and downregulation of a neuronal differentiation marker (elrD) (arrowheads). Abbreviations: n, number of independent experiments; N, number of evaluated embryos in total; Error bars indicate standard error of the means (s.e.m); *p ≤ 0,05; **p ≤ 0,01. P-value were calculated by Student’s T-test. |
