Fig 1: Localisation of GDF5, BMPR1B and BMPR2 in the developing mouse hippocampus. (A) Sections of P10 hippocampus triple labelled with TOTO-3, anti-MAP2 and either anti-GDF5, anti-BMPR1B or anti-BMPR2. Upper panels show low-power images of CA regions and dentate gyrus and the lower panels show high-power images of CA1. Gdf5bp hippocampus was used as a control for anti-GDF5. Controls for anti-BMPR1B and anti-BMPR2 received no primary antibodies. (B) E18 hippocampal neurons double stained with anti-MAP2 and either anti-GDF5, anti-BMPR1B or anti-BMPR2 after 7 days in culture. PL, pyramidal layer; DG, dentate gyrus; sr, stratum radiatum. Scale bars: 200 μm (A, upper panels); 25 μm (A, lower panels; B).
Fig 2: Expression of GDF5 receptors. a, b Expression of transcripts encoding GDF5 receptors in the SCG at stages from E14 to P10 relative to the geometric mean of Gapdh, Sdha and Hprt1 reference mRNAs (mean ± SEM, n = 3 per age). a Relative levels of Bmpr1a and Acvr2a mRNAs normalised to the peak of expression at E14 and P0, respectively. b Relative levels of Bmpr1b and Bmpr2 mRNAs normalised to the peak of expression at E16. c to f Photomicrographs of representative P0 SCG neurons cultured for 24 h with NGF and double labelled for β-III tubulin and either BMPR1A (c), ACVR2A (d), BMPR2 (e) or BMPR1B (f). Scale bar = 40 μm
Fig 3: BMPR1B and BMPR2 are required for GDF5-promoted dendrite growth. (A) Representative E18 neurons transfected after 6 days with plasmids expressing either GFP alone or GFP plus either constitutively active (CA) BMPR1B or dominant-negative (DN) BMPR1B. Six hours after transfection, the neurons were cultured for 18 hours with or without 100 ng/ml GDF5. (B) Percentage of dendrites longer than 50 μm under these experimental conditions. (C) Representative neurons transfected after 6 days with plasmids expressing either GFP alone or GFP plus either wild-type (WT) BMPR2 or dominant-negative BMPR2. After transfection, the neurons were cultured for 18 hours with or without GDF5. (D) Percentage of dendrites longer than 50 μm under these experimental conditions. Mean ± s.e.m. of data from >150 neurons per condition from at least three separate experiments are shown **P<0.001, ***P<0.0001, statistical comparison with control. Scale bars: 50 μm.
Fig 4: Expression of GDF5, BMPR1B and BMPR2 in the developing mouse hippocampus. (A-C) Relative levels of Gdf5 (A), Bmpr1b (B) and Bmpr2 (C) mRNA in the hippocampus (mean ± s.e.m., n=4 per age). (D-F) Representative western blots probed for GAPDH and either GDF5 (D), BMPR1B (E) or BMPR2 (F) and graphs of pro-GDF5, mature GDF5, BMPR1B and BMPR2 relative to GAPDH in the hippocampus (mean ± s.e.m., n=3 per age). Lysates of E18 midbrain were used as positive control.
Fig 5: UMAP Projection of PoPH and non-PoPH cirrhosis tissue and immunohistochemistry of human liver tissue samples.UMAP projection of nuclei derived from PoPH and non-PoPH cirrhosis tissue (A), with contribution of each tissue sample to all identified clusters (B, C). Dot Plot showing differential gene expression of arginine biosynthesis pathway genes (ASL, GPT2, ARG1, GOT1, ASS1, CPS1) in peri-central PoPH clusters relative to non-PoPH cirrhosis clusters. Increased expression of ESR1 (D) and decreased expression of BMPR2 (E) are noted in the region surrounding the central vein in PoPH (right-most column, POL1-POL3) as compared to non-PoPH cirrhosis (middle column, NPOL1–NPOL7). This pattern of protein expression corresponds to the pattern of differential gene expression of BMPR2 and ESR1 in PoPH clusters relative to non-PoPH cirrhosis clusters. Normal healthy liver (NL2, NL4) is also presented as an additional comparator. Note that limited tissue samples prevented us from testing BMPR2 and ESR1 in NL3 samples.
Supplier Page from Abcam for Anti-BMPR2 antibody