Fig. 2

Fig. 2

kif5Ba recruits germ plasm RNAs to cleavage furrows and promotes germ cell specification. (A,B,E,F) nanos3 (A,B) and vasa (E,F) localize to distal furrows in WT (A,E), but not in Mkif5Ba mutants (B,F), at 4-cell stage. Arrowheads indicate nanos3 and vasa accumulations. [nanos3: n=1/28 for Mkif5Ba^ae11/ae12 (N=2 females), n=0/15 for Mkif5Ba^ae11/ae11 (N=1 female), n=30/36 for WT (N=2 females) embryos have signal at ≥1 furrow; vasa: n=0/17 for Mkif5Ba^ae11/ae12 (N=1 female), n=0/19 for Mkif5Ba^ae11/ae11 (N=1 female), n=0/12 for Mkif5Ba^ae12/ae12 (N=1 female), n=31/31 for WT (N=2 females) embryos have signal at e1 furrow.] (C,D,G,H) nanos3 (C,D) and vasa (G,H) are enriched in PGCs in WT (C,G), but not in Mkif5Ba mutants (D,H) at shield stage. Arrowheads indicate PGCs. [nanos3: n=0/32 for Mkif5Ba^ae11/ae12 (N=2 females), n=0/19 for Mkif5Ba^e1/e1 (N=1 female), n=28/28 for WT (N=2 females) embryos have PGCs; vasa: n=0/18 for Mkif5Ba^ae11/ae12 (N=1 female), n=0/22 for Mkif5Ba^ae11/ae11 (N=1 female), n=0/17 for Mkif5Ba^ae12/ae12 (N=1 female), n=37/37 for WT (N=2 females) embryos have PGCs.] (I,J) Vegetal localization of dazl is intact in WT (I) and Mkif5Ba mutants (J) at 4-cell stage. [n=19/19 for Mkif5Ba^ae11/ae12 (N=2 females), n=20/20 for WT (N=2 females) embryos have vegetal signal.] (K) RT-PCR shows that germ plasm RNAs vasa, nanos3 and dazl are inherited by WT and Mkif5Ba mutant eggs (4-cell stage) and are maintained in WT and Mkif5Ba embryos (2hpf). ef1α, loading control. NT, no template. (L-O) Vasa⁺ PGCs are present in WT (L,M) but not in Mkif5Ba mutant (N,O) embryos at 30hpf. Scale bars: 10µm. [n=0/48 for Mkif5Ba^ae11/ae12 (N=2 females), n=19/67 for Mkif5Ba^ae11/ae11 (N=3 females), n=0/20 for Mkif5Ba^ae12/ae12 (N=3 females), n=53/53 for WT (N=4 females) have PGCs.] (P-Q3) H&E-stained transverse sections of adult WT (P) and MZkif5Ba mutants (Q) show that MZkif5Ba testes (green arrows) lack germ cells. P′,Q′ and P′′,Q′′ are higher magnification images of left and right testes, respectively. [n=4/4 Mkif5Ba^ae12/ae12 adult males had empty testes; in addition, n=18/18 adult Mkif5Ba^ae11/ae12 males dissected had testes devoid of germ cells.]