Figure 5

Inhibition of canonical WNT signaling partially rescued the Hmga2 loss-of-function phenotype. (A) Embryonic lung explants were cultured as in Additional file 4: Figure S4A. Explants were treated with control (siCtrl) or Hmga2-specific siRNAs (siHmga2) together with the canonical WNT inhibitor DKK1 for 72 h till stage E15 equivalent (E15*). Scale bars, 500 μm. (B) Terminal branches (top) and total branch length (bottom) were quantified at E12.5 and E15.5* Embryonic lung explants were treated as in A. Error bars show the SEM (n = 6). ***P <0.001; **P <0.01; *P <0.05 after one-way analysis of variance. (C) Inhibition of canonical WNT signaling antagonized the effect of Hmga2-LOF. Protein extracts from the lung explants treated as in A were analyzed by western blot using the indicated antibodies. LMNB1 was used as loading control. (D) Expression analysis of the indicated genes by quantitative RT-PCR in embryonic lung explants that were treated as in A. Relative expression normalized to Tuba1a. Error bars show the SEM (n = 3). Asterisks as B. (E) Protein extracts from the lung explants treated as in A were analyzed by western blot using the indicated antibodies. TUBA1A was used as loading control.