Shh Signaling from the Nucleus Pulposus Is Required for the Postnatal Growth and Differentiation of the Mouse Intervertebral Disc
Figure 4
The effects of Shh targeting in vitro and in vivo on the expression of molecular markers in the AF.
The panels are arranged as in a table. Each vertical column shows a treatment (two or five day vehicle treated control [P4 t2 or 5 Veh] and two or five days in cyclopamine [P4 t2 or 5 CycA] from the in vitro experiments; and control Shh flox [P5 d5 Shhflx/flx; R26rtTA] and Shh mutant [P5 d5; Shhd/d(tetO)7-Cre; R26rtTA] mice from the in vivo experiments). The rows shows the expression, by immunocytochemistry, or Gli1 (A) and (B), Ptch1 (C) and (D), Sox9 (E) and (F), collagen 1 (G) and (H), collagen 2 (I) and (J), chondroitin sulfate (K) and (L), and keratin sulfate (M) and (N). The expression of all these markers was reduced after treatment with cyclopamine in vitro, or Shh targeting in vivo. Scale bars indicate magnifications used. IAF = inner annulus fibrosus, OAF = outer annulus fibrosus. Blue = cell nuclei stained with POPO-3, green = general counterstain with wheat germ agglutinin. See text for details.