Twisted Gastrulation, a BMP Antagonist, Exacerbates Podocyte Injury
Figure 1
Expression and function of Bmps and Bmp antagonists in cultured podocytes.
(A–C) Murine podocytes cultured under non-permissive conditions were subjected to quantitative real-time PCR with various primers for Bmps, Bmp receptors and Bmp antagonists using vector control (see method)(n = 3). (A) Expression of Bmps in cultured podocytes. The expression of each Bmp normalized by vector control was shown relative to that of Bmp2. (B) Expression of Bmp receptors in cultured podocytes. Expression of Bmp receptor type Ib and Bmp receptor type II normalized by vector control was shown relative to that of Bmp receptor type Ia. (C) Expression of Bmp antagonists in cultured podocytes. The expression of Bmp antagonists normalized by vector control was shown relative to that of Twsg1. (D–E) The effects of Bmps and Twsg1 administration on cultured podocytes. Podocytes cultured under non-permissive conditions were stimulated with Bmp and Twsg1, and incubated for an additional 14 days. Recombinant Bmp7 (300 ng/ml), Bmp4 (300 ng/ml), Twsg1 (300 ng/ml) were added to the cultured podocytes at day 1 and day 7. (D) Morphological changes in podocytes after the administration of Bmp4, Bmp7 and Twsg1. Surface area (pixel) of each podocyte was analyzed (n = 30 in each group). (E) The expression of podocalyxin in podocytes treated with Bmp4, Bmp7, and Twsg1. The results of quantitative real-time PCR analysis (n = 7) and representative immunostaining were shown. (F) Proliferation assay of podocyte in the presence of Bmp and Twsg1. We utilized recombinant Bmp7 (300 ng/ml), Bmp4 (300 ng/ml), and Twsg1 (300 ng/ml). (n = 11 in Bmp7 experiment, and n = 8 in Bmp4 experiment) Data are presented as means ± SD. *p<0.05, ***p<0.001; NS, no significant difference. Scale bars: 100 µm.