goat anti jag1  (R&D Systems)

Journal: bioRxiv

Article Title: Single-Cell Spatial Mapping of Human Kidney Development Reveals the Critical Role of the Local Microenvironment in Cell Fate Decisions

doi: 10.1101/2025.06.15.659579

Figure Lengend Snippet: ( a ) Integrated UMAP of spatial (CosMx) and dissociated scRNAseq data showing cell type annotations indicated by color. ( b ) H&E-stained human fetal kidney section adjacent to the corresponding cell type annotation from our spatial transcriptomic data. Cell type annotation is indicated by color. ( c ) Expression of Wilms tumor antigen 1: WT1 , podocin: NPHS2 (podocyte marker genes in yellow) in our analyzed kidney sample. ( d ) Expression of Meis homeobox 2: MEIS2 , UNC homeobox: UNCX , LIM homeobox 1: LHX1 , Apolipoprotein E: APOE , Jagged canonical Notch ligand 1: JAG1 , (yellow) in the developing human kidney. ( e ) Pseudotime of NPC lineages, colored from early (purple) to late (yellow); absorption probabilities for PT, DCT and LOH colored from low (purple) to high (yellow); Renal corpuscle fate absorption probabilities colored from low (blue) to high (red), within the developing human kidney. PT: proximal tubule, DCT: distal convolutes tubule, LOH: loop of Henle, RC: renal corpuscle cells.

Article Snippet: After fixation, explants were blocked in 5% donkey serum and then stained with the following primary antibodies prior to imaging, JAG1 - goat (1:200), (R&D, AF599-SP), Six2 - mouse (1:100) (proteintech, 66347-1-Ig) Six2 – rabbit (1:200) (proteintech 11562-1), LTL (1:200) (Vector Laboratories B-1325), ECAD (1:200) (abcam, ab11512), NPHS1- (R&D, AF4269) sheep (1:200), ITGA8 (1:200) (R&D, AF4076).

Techniques: Staining, Expressing, Wilms Tumor Assay, Marker

Journal: bioRxiv

Article Title: Single-Cell Spatial Mapping of Human Kidney Development Reveals the Critical Role of the Local Microenvironment in Cell Fate Decisions

doi: 10.1101/2025.06.15.659579

Figure Lengend Snippet: ( a ) Spatial locations of CytoSignal ligand scores for IGF1 and IGF2 in the human kidney, with score indicated by color (low being purple and high being yellow). ( b ) Histogram of correlations of microenvironment score with absorption probability for 878 microenvironment signals for the renewal trajectory. X-axis shows correlations and y-axis shows frequency (density estimate) of that correlation. Some individual genes of interest are labeled. ( c ) Ligand score (y-axis) is plotted against pseudotime (x-axis) for IGF2. Each trajectory is shown by its discrete color. ( d ) UMAP of scRNAseq showing That IGF2 is most expressed in the NPC cell population, with cognate receptors IGF1R and INSR expressed in all cell populations. ( e ) EpCam immunofluorescence staining (white: ureteric tree) images and quantified number of UB tips (y-axis) for mouse fetal explants incubated with ectopic IGF2 ligand. Total number of tips was normalized for average number of tips in the control samples for each replicate. This was performed in 3 separate litters. Scale bar = 400 um.} ( f ) EpCam immunofluorescence staining (white: ureteric tree) images and quantified number of UB tips (y-axis) for mouse fetal explants incubated with ectopic insulin ligand. Scale bar = 400 um. This experiment was repeated in a separate mouse litter. Total number of tips was normalized for average number of tips in the control samples for each replicate. This was performed in 3 separate litters. Scale bar = 400 um. Each replicate shown separately in Supplementary Fig. 15. ( g ) SIX2 immunofluorescence staining (white: NPC marker) images and quantified size of SIX2 caps. Area was normalized to average area for control explants for each 3 kidneys per condition performed in 2 separate litters (12 total kidneys). Scale bar = 400 um. Each replicate shown separately in Supplementary Fig. 15. ( h ) SIX2 (purple) and EpCam (white) immunofluorescence of explants treated with IGF1R inhibitor linsitinib at various concentrations. This was repeated in a separate litter and had similar results. Scale bar = 400 um. Each replicate shown separately in Supplementary Fig. 15. ( i ) Phase, SIX2 and JAG1 immunofluorescence of human kidney organoids treated with IGF1R inhibitor linsitinib at various concentrations. This experiment was repeated with similar results in a separate organoid batch. Scale bar indicates 1000 um. ( i ) Histogram of correlations of microenvironment score with absorption probability for 878 microenvironment signals for the PT trajectory. X-axis shows correlations and y-axis shows frequency (density estimate) of that correlation. Some individual genes of interest are labeled. ( j ) Spatial locations of CytoSignal ligand scores for AVP in the human kidney, with score indicated by color (low being purple and high being yellow). ( k ) UMAP of scRNAseq showing That AVP is most expressed in the PT cell population, with cognate receptors AVPR1A and AVP2R expressed in the DCT cell population. ( l ) Ligand score (y-axis) is plotted against pseudotime (x-axis) for AVP. DCT and PT trajectory are shown by their own discrete color. ( o ) CosMx cell type annotations of renal corpuscles, with PT, podocyte and PEC annotations shown in space. ( p ) Immunofluorescence of human fetal kidney tissue showing ACE2 expression (purple), WT1 expression (green) and DAPI (white) ( q ) UMAP showing probability of trajectories for renal corpuscle and tubular fate indicated for each cell based on color (left). Histogram of ligand correlations with renal corpuscle fate within the PT population. X-axis shows correlations and y-axis shows frequency (density estimate) of that correlation. Individual genes of interest are labeled. Scale bar = 100 uM. ( r ) Spatial locations of CytoSignal ligand scores for AVP in the human kidney, with score indicated by color (low being purple and high being yellow). ( s ) UMAP of scRNAseq showing That AGT, and ACE2 are expressed in the PT cell population, with cognate receptor AGTR2 expressed in all NPC derived cell types. PT: proximal tubule, LOH: loop of Henle, DCT: distal convoluted tubule, PEC: parietal epithelial cell, NPC: Nephron progenitor cell, Int: intermediate. IGF2: insulin like growth factor 2, AVP: arginine vasopressin, IGF1: insulin like growth factor 2. AVP: Arginine vasopressin. AGT: Angiotensinogen, ACE2: Angiotensin converting enzyme 2, AGT2R: Angiotensin 2 receptor, AVPR1A: arginine vasopressin receptor 1a, AVPR2: arginine vasopressin receptor 2. MsE: mouse explant. HumSpatial: Human Spatial, HumOrg: Human organoids, HumIF: human immunofluorescence.

Article Snippet: After fixation, explants were blocked in 5% donkey serum and then stained with the following primary antibodies prior to imaging, JAG1 - goat (1:200), (R&D, AF599-SP), Six2 - mouse (1:100) (proteintech, 66347-1-Ig) Six2 – rabbit (1:200) (proteintech 11562-1), LTL (1:200) (Vector Laboratories B-1325), ECAD (1:200) (abcam, ab11512), NPHS1- (R&D, AF4269) sheep (1:200), ITGA8 (1:200) (R&D, AF4076).

Techniques: Labeling, Immunofluorescence, Staining, Incubation, Control, Marker, Expressing, Derivative Assay

Journal: iScience

Article Title: A developmental atlas of mouse vestibular-like inner ear organoids

doi: 10.1016/j.isci.2025.111817

Figure Lengend Snippet:

Article Snippet: goat anti-Jag1 , , Santa Cruz , RRID: AB_649689.

Techniques: Generated, Recombinant, Software

Journal: bioRxiv

Article Title: EBF1 limits the numbers of cochlear hair and supporting cells and forms the scala tympani and spiral limbus during the development of the inner ear

doi: 10.1101/2023.04.28.538789

Figure Lengend Snippet: A. in situ hybridization of Fgf10 and Bmp4 and immunostaining of JAG1 (green) on cross sections of E13.5 and E14.5 Ebf1 +/+ and Ebf1 -/- mice cochlear basal-to-middle region. Fgf10 and Bmp4 expression in the Ebf1 -/- mice cochlea was similar to that in Ebf1 +/+ mice cochlea. In contrast, JAG1 expression area of the Ebf1 -/- mice cochlea was expanded toward the more medial side (A, arrowheads) compared with that of Ebf1 +/+ mice cochlea. The prosensory domain is shown in brackets. B . in situ hybridization for Atoh1 on cross sections of E14.5 and E15.5 Ebf1 +/+ and Ebf1 -/- mice cochlear basal-to-middle region. In the inner ear of E14.5, Atoh1 expression was observed in the cochlea and vestibule in Ebf1 +/+ mice, while in Ebf1 -/- mice, Atoh1 expression was observed in the vestibule, but not in the cochlea. In the cochlea of E15.5, Atoh1 was expressed in both Ebf1 +/+ and Ebf1 -/- mice. Areas enclosed by dashed lines indicate cochlear ducts and vestibules. Scale bars: 100μm.

Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-EBF1 antibody (1:1000, Millipore, Darmstadt, Germany, AB10523), mouse anti-MYO7A antibody (1:1000, Developmental Studies Hybridoma Bank[DSHB], Iowa City, IA, USA, 138-1), rabbit anti-MYO7A antibody (1:1000, Proteus BioSciences, Waltham, MA, USA, 25-6790), goat anti-SOX2 antibody (1:250, R&D Systems, Minneapolis, MN, USA, AF2018), rabbit anti-SOX2 antibody (1:100, Proteintech, Manchester, UK, 11064-1-AP), rabbit anti-VGLUT3 antibody (1:500, Synaptic systems, Goettingen, Germany, 135 203), goat anti-JAG1 antibody (1:500, Santa cruz, Dallas, Texas, USA, sc-6011), mouse anti-p27Kip1 antibody (1:200, BD Transduction Laboratories, 610242), rabbit anti-Tubulin β 3 (TUJ1) antibody (1:1000, Biolegend, San Diego, CA, USA, PRB-435P), rabbit anti-PROX1 antibody (1:500, Millipore, AB5475), rabbit anti-p75 antibody (1:500, Millipore, AB1554), and rabbit anti Cleaved caspase3 (CC3) antibody (1:400, Cell Signaling Technology, Danvers, MA, USA, Asp175).

Techniques: In Situ Hybridization, Immunostaining, Expressing