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Fig. 1. Mlc1v-nLacZ-24+/– expression recapitulates
Fgf10 expression. (A) Fgf10 expression by whole-mount
in-situ hybridization in E10.5 wild-type control lungs. Note the expression in
the right distal lung mesenchyme (black arrows), the thyroid and the
developing stomach. (B) X-gal-stained E10.5
Mlc1v-nLacZ-24+/– lungs recapitulate the
Fgf10 expression pattern at this stage in the distal lung mesenchyme
(black arrow) and the thyroid. (C) Fgf10 expression by whole-mount
in-situ hybridization in E11.5 wild-type control lungs. Note the expression in
the left distal lung mesenchyme (white dotted box). (D) X-gal staining of
E11.5 Mlc1v-nLacZ-24+/– lungs recapitulates the
Fgf10 expression pattern at this stage, except in the left lobe
(white dotted box). (E) Fgf10 expression by whole-mount in-situ
hybridization in E12.5 wild-type control lungs. Note the expression in the
distal lung mesenchyme (black arrows). The area in the dotted box is magnified
in (F). (G) LacZ expression at RNA level in
Mlc1v-nLacZ-24+/– lungs. (H) LacZ is
expressed at the tip of the accessory lobe recapitulating the Fgf10
pattern (dotted box in G). Notice the absence of Fgf10 expression
close to the epithelium (small double white arrow) (I) ß-galactosidase
activity shown by X-gal staining is found in the distal mesenchyme (white
arrow) and at the level of the bronchi of
Mlc1v-nLacZ-24+/– lungs (small white arrows). Note
that X-gal staining is now present in the mesenchyme of the left lobe. Note
also that ß-galactosidase-positive cells are not detected in the primary
bronchi (black arrows). (J) High magnification of the accessory lobe (dotted
box in I). (K) Fgf10 expression at RNA level by whole-mount in-situ
hybridization in E14.5 wild-type control lungs. Note the expression in the
mesenchyme at the periphery of the lobes. (L) X-gal staining of E14.5
Mlc1v-nLacZ-24+/– lungs showing LacZ
expression at the periphery of the lobes similar to the Fgf10
expression pattern. Note ß-gal expression at the level of the bronchi
(black arrow). (M) High magnification of the surface of the cranial lobe shown
in L. (N-Q) Control E11.5 Mlc1v-nLacZ-24+/– lung
grown in absence of cyclopamine. (O) After 28 hours in culture new branches
are formed. Note that the distal epithelium is not dilated (arrow). (P) X-gal
staining of the cultured lung shown in O. ß-gal expression is found in
the distal mesenchyme. (Q) Vibratome section through the left lobe shown by
the arrow in P. Note the low level of ß-gal expression. (R-U) E11.5
Mlc1v-nLacZ-24+/– lung grown in presence of 5
µmol/l of cyclopamine. (S) After 28 hours of culture the lung exhibits
dilated end buds (arrow). (T) X-gal staining of the cultured lung shown in S.
Note the increase in ß-gal expression throughout the lung in comparison
with the lung grown in the absence of cyclopamine shown in P. (U) Vibratome
section through the left lobe of the lung shown by the arrow in T. Note the
marked increase in LacZ expression compared with the untreated lung
(Q). Scale bar: 110 µm in A,B; 180 µm in C,D; 210 µm in E,G,I; 105
µm in F,H,J; 435 µm in K,L; 80 µm in M; 175 µm in J; 250 µm in
N,O; 300 µm in O,S; 190 µm in P,T; 50 µm in Q,U. acc, accessory lobe;
br, bronchus; cont, control; cran, cranial lobe; st, stomach; th, thyroid; tr,
trachea.
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