Fig. 4. Surfactant expression is reduced during embryogenesis in DKO mice and is regulated by HMGA1 and HIPK2 expression in cells.

a RNA extracted from lungs of WT and DKO mice at E17.5 d.p.c. was analyzed by qRT-PCR for Sp-A, Sp-B, and Sp-C expression. The Actin expression level has been used for normalization. Data are mean ± SD of a representative experiment performed in triplicate; * indicates significant difference P < 0.01, ***P < 0.001. n = 3 WT mice and n = 10 DKO mice were analyzed. b Immunohistochemistry for SP-C on WT and DKO embryos (× 200 magnification). One representative picture is shown. c HeLa cells were transfected with Nkx2.1 expression vector alone (Ctrl) or with the indicated plasmids. All transfections were performed in triplicate. Data are mean ± SD of a representative experiment performed in triplicate; * indicates significant difference P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test). d BEAS-2B cells were transfected with control (CN), anti-HMGA1 (A1i), anti-HIPK2 (K2i), or anti-HMGA1 and anti-HIPK2 (A1i + K2i), and, after 72 h, proteins were extracted to perform western blot analysis with the indicated antibodies. Anti-actin and anti-vinculin antibodies were used as loading control