Journal: The EMBO Journal

Article Title: BMP4 initiates and patterns ventral-caudal structures in zebrafish and human pluripotent stem cell aggregates

doi: 10.1038/s44318-025-00643-6

Figure Lengend Snippet: ( A ) Representative images of bmp4 -injected zebrafish embryos (upper) and explants (lower, with injection site towards the right) from 2.25 to 24 hpf. Red fluorescence marks the descendants of the bmp4 -injected blastomere. Arrowheads at 8 hpf and 10 hpf embryo point to the protrusions induced by Bmp4 in embryos, and at 24 hpf in explants to the segmented somites. Labels indicate the primary (i) and secondary axis (ii). ( B ) Immunofluorescence imaging of Phospho-Smad1/5/9 in bmp4 + RFP injected explant (upper) and RFP injected explant (lower) at 4 hpf, with the injection site towards the right. Signal measurements along the dotted rectangles are shown on the right. Dark Green curves represent the nonlinear fit of actual data curves (light green), with Dark Green numbers indicating the mean slopes. Red curves depict the distributions of RFP signaling intensity along the dotted rectangles. ( C ) Quantification of the elongation of Bmp4 explants from 4 to 24 hpf. The Midlines along anterior–posterior axis of Bmp4 explants are measured (red line), as displayed in the upper left. Data are presented as mean ± SD. ( D ) bmp4 -injected explants (upper, posterior towards the right) and embryos (lower, animal pole view) at 6 hpf stained by whole mount in situ hybridization (WISH) with the indicated probes. DAB staining of GFP (brown spots) indicates the clones of bmp4 & GFP mRNA injected blastomere at 2.25 hpf. ( E ) Bmp4 explants at 14 hpf stained by WISH with indicated probes. ( F ) Whole mount in situ hybridization chain reaction (HCR) staining of Bmp4 explants at 12 hpf for tbxta and sox2 , with co-staining of DAPI. NT neural tube, MP neuromesodermal progenitor, TMP tailbud mesodermal progenitor. ( G ) Time-lapse imaging of Bmp4 explants displaying the sequential formation of somites, as per Movie . White dotted lines indicate existing somites, and purple dotted lines indicate newly formed somites. Nuclei were labeled by H2B-GFP (green fluorescence). ( H ) Bmp4 explants at 24 hpf stained by WISH with the indicated probes. For all explants displayed in this article, anterior (opposite to the injection site) is toward the top unless otherwise stated. Each experiment was performed for at least three independent replicates (technical replicates). Scale bars: 100 μm. .

Article Snippet: HCR probe wash buffer , Molecular Instruments , BPW02424.

Techniques: Injection, Fluorescence, Immunofluorescence, Imaging, Staining, In Situ Hybridization, Clone Assay, Labeling

Journal: Developmental biology

Article Title: PRDM paralogs are required for Meckel’s cartilage formation during mandibular bone development

doi: 10.1016/j.ydbio.2025.12.002

Figure Lengend Snippet: (A,B-A””,B””) Hybridization chain reaction (HCR) was performed on coronal sections of E10.5 mandibular processes to assess gene expression patterns for Dlx5 (green) (A-B), Dlx6 (cyan) (A’-B’), Hand2 (magenta) (A”-B”), Sox10 (yellow) (A”’-B”’) in controls and double mutants. (A””-B””) Merged image of A-A”’ and B-B”’ with the counterstain DAPI in blue. (C) RT-qPCR for the indicated mRNA transcripts isolated from the mandibular processes of control and Prdm3;Prdm16 double mutants at E10.5 (n = 4 for both control and mutants). (D) RT-qPCR for Sox10 at E10.5 (n = 4 for both control and mutants) and E11.5 (n = 3 for control, n = 4 for mutants). (E-F, E’-F’) HCR was performed on coronal sections of E11.5 mandibular processes to assess gene expression patterns for Sox10 . (E’-F’) Merged image of E and F with the counterstain DAPI in blue. Double white arrowhead denotes the initial presumptive cranial ganglia and peripheral nerve tracks entering the mandibular prominence in control animals. White arrow indicates areas of Sox10 expression that are expanded in the mesenchyme in the Prdm3;Prdm16 double mutants. Abbreviations: MdP, mandibular process. *P≤0.05; (unpaired, two-tailed Student’s t test).

Article Snippet: Sections were pre-hybridized in HCR HiFi Probe Hybridization Buffer (Molecular Instruments) for 10 minutes at 37°C in a humidified chamber.

Techniques: Expressing, Hybridization, Gene Expression, Quantitative RT-PCR, Isolation, Control, Two Tailed Test