Fig. 6. Forced Pitx2c expression in posterior SHF explants from
Mlc3f-nlacZ-2E embryos at the 5-somite stage. (A-H)
Control GFP adenovirus infection on left (L) (A-D) and right (R)
(E-H) explants from the pSHF of Mlc3f-nlacZ-2E mouse embryos. (A,E)
Merge of phase-contrast and GFP fluorescence on the live explants; (B,F) GFP
fluorescence on the live explants; (C,G) Hoechst staining; (D,H) β-gal
immunofluorescence on explants now spread under a coverslip. (I-P)
Pitx2c/GFP adenovirus infection on left (L) (I-L) and right (R) (M-P)
explants of the pSHF from Mlc3f-nlacZ-2E embryos. (I,M) Merge of
phase-contrast and GFP fluorescence on live explants; (J,N) GFP fluorescence
on the live explants; (K,O) Hoechst staining; (L,P) β-gal
immunofluorescence on explants now spread under a coverslip. Scale bar: 10
µm in P for A-P. (Q) Quantitative analysis of the percentage of
β-gal-positive cells in the total cell population, after infection with
the control GFP adenovirus (white and black; P
0.025) or
with the Pitx2c/GFP-expressing adenovirus (grey and dark grey). The
P-value for left-GFP compared with left-Pitx2c is 0.01 and for
right GFP compared with right Pitx2c is 0.005. White and grey, left
explants; black and dark grey, right explants. The quantification was
performed on ten explants for each side and for each adenovirus. (R)
qRT-PCR analysis after GFP and Pitx2c/GFP infection on five
pooled left explants (l) and five pooled right explants (r) after 72 hours of
culture for each adenovirus. lacZ (arrowheads point to the difference
in rlacZ), GFP and Mlc2a transcript abundance is
expressed as a percentage of that of Gapdh transcripts.