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Fig. 2. Knockdown of xKIF13B results in PGC mislocalization. All embryos were stained for Xpat (PGC marker) and MyoD (somite marker). (A and B) Representative examples of embryos injected with 33.6 ng of control MO (A), or with 16.8 ng of xKIF13B M02 (B). Quantifications of the average number of mislocalized PGCs (C and E) and of PGC “spreading” (D and F) in control MO (C and D) and xKIF13B MO injected (E and F) embryos (see also Material and methods). (G) xKIF13B loss- and gain-of-function reduces the average PGC number and leads to PGC mislocalization. Control embryos were injected with 33.6 ng of control MO: xKIF13B morphants were injected with 16.8 ng of xKIF13B MO2; rescue was performed by co-injection of 16.8 ng of xKIF13B MO2 and 0.1 ng of the xKIF13B ORF_DE mRNA; for xKIF13B overexpression 0.1 or 1 ng of the xKIF13B ORF_DE mRNA per embryo was injected. For each independent injection 20 embryos per treatment were analyzed. Average PGC numbers were calculated in percentage relative to the control embryos set to 100% for each independent round of injections. PGC spreading represents the minimal ellipse covering the germ cells, normalized to the size of the embryo and represented in arbitrary units (a.u). PGC mislocalization was calculated as percentage of mislocalized (ectopic) PGCs relative to the total PGC number. PGCs were regarded as “mislocalized” when positioned beyond somites 5–11 along the A/P axis of an embryo. Results were averaged between at least three independent rounds of injections. Error bars represent the standard deviation of the mean, lines depict p < 0.05. |
