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Fig. 5. High levels of dpERK activity in somites results in loss of MyoD
and Mkp3. (A) In situ hybridisation for Mkp3 after
electroporation of a plasmid encoding caMEK1, a constitutively active form of
MKK1; arrow indicates ectopic Mkp3 transcripts in the neural tube,
bracket indicates the loss of Mkp3 transcripts in targeted somites
(n=30/35). (B) Western blot detecting dpERK in somites electroporated
with caMEK1 plasmid (+) compared with control somites (-); 7.5 µg of
protein was loaded in each lane, compared with twice that amount in
Fig. 3D and
Fig. 4A. (D-G) In situ
hybridisation for MyoD. (D) hMKP3-GFP electroporation has no effect
on MyoD expression (n=16/16). INT/BCIP was used to detect
GFP probe (red), it was washed out using methanol to visualise MyoD
(E). (F) caMEK1 electroporation caused complete loss of MyoD
transcripts (blue, indicated by bracket; n=10/15) but Pax3
(C) was not affected (n=16/16). (G) FGF8 beads (asterisk) resulted in
loss of MyoD in adjacent somites (n=7/11). (H) caMEK1
electroporation caused a significant loss of Fgf4 transcripts (blue,
indicated by bracket; n=13/18).
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