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Fig. 8. Interfering with ephrin A/EphA signalling in ovo led to inappropriate
caudal turning of some cC-VC axons. RCASB vector or RCASB-EphA3
C
was electroporated in ovo into caudal hindbrain at stage 20-21. An EGFP
expression vector was co-electroporated to reveal the domain of successful
electroporation. At stage 27-28, electroporated hindbrains were taken for
culture and cC-VC axons were anterogradely labelled. (A-D) RCASB vector
was electroporated. (E-H) RCASB-EphA3C was electroporated. EGFP
signals in A,E indicate the domain of electroporation. (B,F) The same images
as in A,E, respectively, with EGFP signals removed to better reveal the axons
at their turning point. (C,D,G,H) Higher magnifications of axon turning
regions of A,B,E,F, respectively. (I) A different sample with a higher
degree of caudal-turning error when RCASB-EphA3C was electroporated.
(J) EphA3-AP in situ binding on a hindbrain co-electroporated with
RCASB-EphA3C and EGFP. The electroporated domain of this sample
indicated by EGFP is outlined with white dots. (K) Quantification of
caudal turning error in hindbrains electroporated with RCASB-EphA3C
versus RCASB mock vector (Control). The degree of caudal turning is
represented as the ratio of caudal-over rostral-turning axons. Data are
represented in a scatter chart, with the sample values sorted in an ascending
order. The red and orange arrows indicate the data points of samples shown in
E-H and in I, respectively. Arrows in F,H,I indicate axons turning caudally.
Samples with RCASB-EphA3C showed significantly higher incidence and
amount of inappropriate caudal turning (P<0.0001, Mann-Whitney
U-test). Scale bar: 200 µm in A,B,E,F,I,J; 100 µm in
C,D,G,H.
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