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Fig. 5. Electroporation of
Ptc1
loop2 effectively blocks
Shh signal transduction in the central retina. (A-C) Coronal
sections of E16.5 retinas electroporated with EGFP and
Ptc1
loop2 (B,C) or EGFP alone (A)
and immunostained with anti-Isl2 (C-RGCs) or anti-Zic2 (I-RGCs, arrows).
(D) Quantitative RT-PCR analysis of Ptc1, Ptc2 and
Hip1 mRNA levels. Values were normalised to 18S rRNA. In
Ptc1
loop2-electroporated retinas, the
endogenous levels of Ptc1 (Ptc1 3'UTR), Ptc2 and
Hip1 mRNA are significantly reduced. Primers recognising the
Ptc1
loop2 form (Ptc1) confirmed effective
plasmid transduction. Values represent the mean ± s.e.m. of two
independent experiments performed in triplicate.
*P<0.05,**P<0.01 (Student's
unpaired t-test). Scale bar: 200 µm.