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Fig. 5. Gene silencing of TSK results in the inhibition of organizer
formation. (A) Transplantation scheme of siRNA experiments. Two
chick embryos were electroporated with siRNA at stage 3+ and the
excised MPSs were implanted into the lateral Hensen's node of the host embryo.
(B) The embryo at stage 3+ was electroporated with control1,
TSKA or TSKs siRNA. Hensen's node and the anterior fourth of the
primitive streak were surgically removed. After 6 hours incubation, the level
of VG1 or TSKB mRNA in the MPS was determined by RT-PCR.
(C-J) After carrying out experiments according to the diagram in A, the
ectopic induction of chordin (C-F) or goosecoid (G-J) was examined by in situ
hybridization. (D,E) In the case of control1 and TSKA siRNAs, the MPS
induced ectopic expression of chordin in 17/44 (39%) and 12/32 (38%),
respectively, which are similar to embryos without electroporation (C) [20/50
(40%)]. (F) In the case of TSKs siRNA, the MPS induced ectopic expression of
chordin in 8/40 (20%) embryos. (H,I) In the case of control1 and TSKA
siRNAs, the MPS induced ectopic expression of goosecoid in 12/42 (29%) and
12/44 (27%), respectively, which are similar to embryos without
electroporation (G) [12/40 (30%)]. (J) In the case of TSKs siRNA, the MPS
induced ectopic expression of chordin in 7/53 (13%) embryos.
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