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 ) Immunofluorescence staining for phosphorylated Smad1/5/9 in embryos injected with bmp4 + RFP mRNA (left) and RFP mRNA alone (right) at 6 hpf. The injection site was determined by red fluorescence. Signal quantifications along the ventro-dorsal axis are displayed beside the merged images. ( B ) Immunofluorescence staining for pSmad1/5/9 in explants injected with bmp4 + RFP mRNA (left) and RFP mRNA alone (right) at 10 hpf. Signal quantifications along the anterior–posterior axis are shown next to the merged images. Clones of bmp4 injected blastomere would stay at the injected end when they differentiated into enveloping layer cells. But when they differentiate into deep cells, they would involute and migrate anteriorly. Signal intensities of pSmad1/5/9 and RFP were normalized first by the DAPI signal and then by their mean signal intensity. ( C ) Whole mount in situ hybridization (WISH) of id1 (left) HCR co-staining of id1 , tbxta and tbx6 (middle) in Bmp4 explants at 14-18 hpf, with injection sites oriented to the right. Signal quantifications along the white dashed line are presented to the right. ( D ) WISH of eve1, fgf8a, sox32, cdx4, chrd in Bmp4 explants at 6 hpf, with injection sites oriented to the right. ( E ) WISH of tbxta, cdx4 in Bmp4 explants treated with Nodal inhibitor (SB505124, 50 μM), Wnt inhibitor (IWP-L6,25 μM) or FGF inhibitor (SU5402, 25 μM) at 6 hpf, with injection sites oriented to the right. Representative images of treated or untreated Bmp4 explants at 24 hpf are displayed. ( F ) WISH of wnt8a, ndr2 , and tbxta in uninjected, 20 pg bmp2b mRNA injected, 0.5 pg bmp4 mRNA injected explants or embryos at 6 hpf. ( G ) Expression patterns of fgf8a , sox17 , and eve1 revealed by WISH in bmp4 -injected embryos at 6 hpf. Views from the animal pole of embryos are displayed, with descendants of the bmp4 -injected blastomere labeled by DAB staining of GFP (left and middle images). ( H ) WISH of sox32 in uninjected, 8 pg or 20 pg bmp4 mRNA injected explants at 10 hpf. ( I ) Expression pattern of tbxta revealed by WISH in bmp4 -injected embryos at 12 hpf. ( J ) WISH for tbxta, egr2b, sox19a, olig2, foxd3, shha, tnnt2c, myod1, cldn3d, hoxc13b in bmp4 -injected embryos 24 hpf. ( K ) WISH for egr2b, gata6, shha, her1, fn1b , and foxd3 in Bmp4 explants at 24 hpf. Each experiment was performed for at least three independent replicates (technical replicates). Scale bars: 100 µm.

Article Snippet: SU5402 , MedChemExpress , HY-10407.

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

Journal: Current Issues in Molecular Biology

Article Title: Pro-Angiogenetic Effects of Purified Extracts from Helix aspersa during Zebrafish Development

doi: 10.3390/cimb44080232

Figure Lengend Snippet: Pretreatment with a VEGF pathway inhibitor prevented the formation of intersomitic vessels: The panels show representative images of a WISH assay performed with fli1 as an endothelial marker, at 36 hpf, in untreated ( A , C , E , G , I ) and treated embryos ( B , D , F , H , J ) with SUGEN 5416. Previously, some embryos were treated with snail derivatives via Method I and controls with fish water and 0.1% DMSO (see Materials and Methods, and ). Magnification of the trunk region (32×). Two replicates were performed ( n = 15). Ratios at the bottom-left part of each picture specify the number of embryos showing the same staining pattern, compared to the total number of embryos used for each experiment. The images were taken in lateral position at 32× magnification with a Zeiss Axiozoom V13 (Zeiss, Jena, Germany) microscope, equipped with a PlanNeoFluar Z 1×/0.25 FWD 56 mm lens and Zen Pro software.

Article Snippet: Zebrafish embryos were then treated at 4 hpf with the snail derivatives LH, LM, LH3, and LM2, and then at 15 hpf with the VEGF inhibitor SU5416 (Sunitinib, Merck KGaA, Darmstadt, Germany) at a final concentration of 5 μg/mL in fish water and incubated in 3 cm diameter well-plates.

Techniques: Marker, Staining, Microscopy, Software