sheep anti 156 vsx2  (Absolute Biotech Inc)

Journal: Genes & Development

Article Title: DIO3 coordinates photoreceptor development timing and fate stability in human retinal organoids

doi: 10.1101/gad.352924.125

Figure Lengend Snippet: DIO3 is expressed in RPCs in the developing human retina and retinal organoids. ( A ) UMAP from CZ CellxGene Discover of human fetal retina scRNA-seq with major cell types labeled. ( B ) DIO3 mRNA expression in fetal retina is limited to DIO3 + RPCs, overlaid on the unannotated UMAP. ( C ) DIO3 is expressed in VSX2 + cells in a 14 week old fetal retina. ( D–F ) DIO3 expression in human retinal organoids at days 80 ( D ), 140 ( E ), and 200 ( F ). ( G ) DIO3 and Ki67 expression in human retinal organoids at day 90. Green circles indicate Ki67 + /DIO3 + cells. ( H ) DIO3 and cone opsin expression in human retinal organoids at day 140. Yellow outlines indicate L/M-opsin + cells. Blue outlines indicate S-opsin + cells. ( H ) Region of morphologically naive photoreceptors within an organoid (same organoid as in I ). ( I ) Region of morphologically more mature photoreceptors within an organoid (same organoid as in H ). ( J ) DIO3 activity as measured by deiodination activity of wild-type organoids over time. Error bars represent SEM.

Article Snippet: Antibodies used in this study were as follows: goat ARR3 (1:200; Novus Biologicals NBP1-37003), mouse CRX (1:500; Abnova UJOHO013), mouse CRX (1:200; Sigma-Aldrich WH0001406M2), rabbit DIO3 (1:200; Novus Biologicals NBP1-05767), mouse Ki67 (1:200; Santa Cruz Biotechnology sc-23900), goat NRL (1:200; Bio-Techne AF2945), rabbit OPN1LW/OPN1MW (1:200; Sigma-Aldrich AB5405), chick OPN1LW/OPN1MW (1:200; kind gift from Jeremy Nathans), goat OPN1SW (1:200; Santa Cruz Biotechnology sc-14363), chick OPN1SW (1:200; kind gift from Jeremy Nathans), rabbit RCVRN (1:200; Sigma-Aldrich AB5585), rat RFP (1:400; Proteintech 5F8-150), mouse rhodopsin (1:500; GeneTex GTX23267), mouse rhodopsin (1:500; Thermo Fisher Scientific MA5-11741), and sheep VSX2 (1:100; Exalpha Biologicals X1180P).

Techniques: Labeling, Expressing, Activity Assay

Journal: iScience

Article Title: The RBPMS CreERT2-tdTomato mouse line for studying retinal and vascular relevant diseases

doi: 10.1016/j.isci.2023.108111

Figure Lengend Snippet: Expression of RBPMS in adult RBPMS CreERT2-tdTomato mouse ocular tissue (A) Representative image of an adult mouse ocular frozen section after tamoxifen induction with strong tdTomato fluorescence signals present in the retinal ganglion layer (RGL), optic nerve, choroid vasculature and ciliary body. Scale bar, 500 μm. (B and C) Ocular frozen sections labeled with RBPMS by immunofluorescent staining (green), and the ratio of tdTomato + and RBPMS + cells. Data are represented as mean ± SEM. Scale bar, 100 μm. (D and E) Ocular frozen sections labeled with Tuj-1 by immunofluorescent staining (green), and the ratio of tdTomato + and Tuj-1 + cells. Data are represented as mean ± SEM. Scale bar, 100 μm. (F and G) Ocular frozen sections labeled with Calretinin by immunofluorescent staining (green), and the ratio of tdTomato + and Calretinin + cells. Data are represented as mean ± SEM. Scale bar, 100 μm. (H) The ocular frozen sections were labeled with tdTomato by immunofluorescent staining (green). The right images show a higher magnification of the corresponding white box area. Scale bar, 100 μm. (I) Ocular frozen sections were labeled with VSX2 by immunofluorescent staining (pseudo color green), Scale bar, 100 μm. (J) The ocular frozen sections were labeled with Recoverin by immunofluorescent staining (green), Scale bar, 100 μm.

Article Snippet: Sheep polyclonal anti-Chx10 (VSX2) , Sigma-Aldrich , Cat# AB9016; RRID: AB_3064860.

Techniques: Expressing, Fluorescence, Labeling, Staining

Journal: iScience

Article Title: The RBPMS CreERT2-tdTomato mouse line for studying retinal and vascular relevant diseases

doi: 10.1016/j.isci.2023.108111

Figure Lengend Snippet:

Article Snippet: Sheep polyclonal anti-Chx10 (VSX2) , Sigma-Aldrich , Cat# AB9016; RRID: AB_3064860.

Techniques: Recombinant, Lysis, Protease Inhibitor, Plasmid Preparation, Software

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: ( A ) A scheme of the procedure. ( B ) Diagrams of developing CONCEPT organoids showing concentric zones of the anterior ectodermal progenitors. A summary of , and , – . ( C ) Morphology of cysts at day 2 showing the epithelial structure indicated by apical localization of the reporter TJ::GFP at the lumen. ( D ) Morphology of CONCEPT organoids at day 26. ( E–G ) Expression of telencephalon (Tel) marker Foxg1, neuroretinal (NR) markers Vsx2 and Pax6 in mouse eyes at E10-10.5. Rostral optic stalk (OS) connected the telencephalic vesicle to the optic cup. ( H–O ) FOXG1+ telencephalic progenitors, VSX2+ and/or PAX6+ retinal progenitors formed concentric zones in CONCEPT organoids. N>5 experiments. ( P–W ) In CONCEPT organoids, morphogens FGF8, BMP4, and BMP7 mRNA expression started at early stages and subsequently formed circular gradients. N>5 experiments. Scale bars, 100 µm ( C, E, M, O, P, S, T ), 200 µm ( I, K ), 500 µm ( Q, U ), 1 mm ( D, H, J, L, N, R, V, W ).

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing, Marker

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: ( A–E ) Images of VSX2 and PAX6 expression ( A ), PAX6 ( B ), VSX2 ( C ), DAPI ( D ), and an image at a higher magnification ( E ) are shown. Scale bars, 1 mm ( A ), 200 µm ( E ).

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: ( A–E ) Images of VSX2 and PAX6 expression ( A ), PAX6 ( B ), VSX2 ( C ), DAPI ( D ), and an image at a higher magnification ( E ) are shown. Scale bars, 1 mm ( A ), 100 µm ( E ).

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: N>5 experiments. ( A–D ) POU4F2+ RGCs grew TUBB3+ axons toward and then along a path with a circular or a portion of circular shape. ( E, F ) In mice, Pax2 was expressed in central regions of the retina and optic stalk at E10.5 ( E ) and in the optic disc and optic stalk at E13.5 ( F ). Tubb3+ axons from the initial RGCs grew toward the optic disc, exited the eye, and navigated within the optic stalk ( G ). ( H–L ) In CONCEPT organoids at day 26, TUBB3+ RGC axons grew toward and then along a path defined by an adjacent PAX2+ VSX2+ cell population (arrowhead in H, brackets in I, J ); the PAX2+ VSX2- cell population set up an inner boundary of RGC axon growth. ( L ) A diagram summarizing RGC axon growth, PAX2+ VSX2+ optic disc (OD), and PAX2+ VSX2- optic stalk (OS) in CONCEPT organoids. The area labeled by the asterisk may appear as false signals in a low-resolution printout but it is clearly a background in digital display. ( M ) A count of CONCEPT organoids showing directional retinal ganglion cell axons. Scale bars, 50 µm ( E, F, G ), 100 µm ( B ), 200 µm ( A, H, I ).

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Labeling

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: CONCEPT organoids at day 24 were used for profiling. ( A ) Identification of 14 cell clusters. ( B ) Cell cycle phases revealed by cell cycle scores. ( C ) FOXG1 expression marked telencephalic cells. ( D, E ) The expression of PAX6 and/or VSX2 marked retinal cells. ( F ) PAX2+ cells were found in two major cell populations: PAX2+ VSX2+ cells were assigned as the optic disc (OD), whereas PAX2+ VSX2- FOXG1+ cells were assigned as the optic stalk (OS). ( G–L ) The expression of major DEGs in cluster 2, the major cell population that mimics the optic disc. ( M–P ) The expression of major gene markers for PAX2+ VSX2- optic stalk cells. ( Q–T ) Identification of CNTN2 as a specific marker for early human RGCs. A large portion of cluster 11 differentially expressed neurogenic retinal progenitor marker ATOH7 and RGC markers POU4F2 and SNCG . The expression of CNTN2 and POU4F2 largely overlapped. ( U–X ) Two small portions of cluster 11 differentially expressed early photoreceptor cell markers ( OTX2 and CRX , U, V ) and amacrine/horizontal cell markers ( TFAP2C and PTF1A, W, X ), respectively.

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing, Marker

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: Related also to . The dataset deposited by Gabriel et al. was used for plotting. ( A–D ) Cell clustering was reproduced ( A, C ). In Gabriel et al.’s organoids on days 30 and 60, PAX2+ cells were extremely rare ( B, D ). Instead of forming cell clusters, a few PAX2+ cells were scattered across datasets. ( E–G ) In Gabriel et al.’s organoids on day 30, FOXG1+ telencephalic progenitors, SIX3+ retinal progenitors, and ATOH7+ neurogenic retinal progenitors were extremely rare; a few positive cells were scattered across the dataset. VSX2 was not found in the dataset of Gabriel et al.’s organoids on day 30; it was filtered out probably due to its extremely low expression.

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet: ( A ) Cell clustering of human fetal retinas HGW9 (GSE138002) identified 21 clusters. ( B–F ) Cluster 18 differentially expressed PAX2, COL9A3, CYP1B1, SEMA5A, and FGF9 , which were top DEGs of cluster 2 in CONCEPT organoids. ( G ) VSX2 expression marked retinal progenitor cells. ( H–M ) Identification of neurogenic retinal progenitor cells ( H ), early photoreceptor cells ( I ), amacrine/horizontal cells ( J, K ), and early RGCs ( L, M ). POU4F2 and CNTN2 were largely co-expressed in early RGCs, consistent with their expression profiles in CONCEPT organoids. ( N–R ) When cells in cluster 18 and retinal progenitors from HGW9 were combined with cells in clusters 2, 4, 5, 7 from CONCEPT organoids (CR24) for Seurat anchor-based clustering, cells in cluster 18 from HGW9 (H18) were grouped with cluster 2 from CONCEPT organoids (C2, assigned optic disc; N ), and these cells expressed both PAX2 and VSX2 (arrowheads in N-R). A small portion of H18 cells were grouped with C4 cells (assigned optic stalk; N ), and these cells expressed PAX2 but not VSX2 (arrows in N-R).

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Expressing

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Western Blot, Labeling, Sequencing, Software

Journal: eLife

Article Title: Self-formation of concentric zones of telencephalic and ocular tissues and directional retinal ganglion cell axons

doi: 10.7554/eLife.87306

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-VSX2 (sheep polyclonal) , Millipore , Cat# AB9016 , IF (1:500).

Techniques: Western Blot, Labeling, Sequencing, Software