Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: MMP-10 immunostaining in 1-week differentiated pellet cultures of IKKα KD vs. matched WT (GL2 control) human OA chondrocytes. Global views of micromass sphere sections (100Xmagnification) are shown on the left; and three random fields at 400Xmagnification, which were submitted to quantitative image analysis appear to the right. Antibody stained images are shown in the right side upper rows (labeled as 1, 2 & 3), with each 400Xfield submitted to analysis by Nikon Imaging Software in the lower rows (1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining of MMP-10 is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: TIMP-3 immunostaining in 1-week differentiated pellet cultures of IKKα KD vs. matched WT (GL2 control) human OA chondrocytes. Global views of micromass sphere sections (100Xmagnification) are shown on the left; and three random fields at 400Xmagnification, which were submitted to quantitative image analysis, appear to the right. Antibody stained images are shown in the right side upper rows (labeled as 1, 2 & 3), with each 400Xfield submitted to analysis by Nikon Imaging Software in the lower rows (1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining of TIMP-3 is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: COL2 immunostaining in 1-week pellet cultures of primary human OA chondrocytes: WT (GL2 control), IKKα KD cells with empty BIN retroviral vector, IKKα KD cells with WT murine IKKα-BIN and IKKα KD cells with kinase-dead murine IKKα(K44M)-BIN. Global views of micromass sphere sections (100Xmagnification) are shown on the left; and three random fields at 400Xmagnification submitted to quantitative image analysis are on the right. Antibody stained images are shown in the right side upper rows (labeled 1, 2 & 3), with each 400Xfield analyzed by Nikon Imaging Software in the lower rows (labeled 1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Retroviral, Plasmid Preparation, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: C1,2C antibody detection of COL2–3/4C neoepitopes in the same 1-week pellet cultures of primary human chondrocytes employed above in . Global views of micromass sphere sections (100Xmagnification) are shown on the left; and three random fields at 400Xmagnification submitted to quantitative image analysis are on the right. Antibody stained images are shown in the right side upper rows (labeled 1, 2 & 3), with each 400Xfield analyzed by Nikon Imaging Software in the lower rows (labeled 1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Staining, Labeling, Imaging, Software, Immunohistochemistry, Control

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: TIMP-3 immunostaining in 1-week pellet cultures of primary human OA chondrocytes: WT (GL2 control), IKKα KD cells with empty BIN retrovector and IKKα KD cells with kinase-dead murine IKKα(K44M)-BIN. Global views of micromass sphere sections (100Xmagnification) are shown on the left, and three random fields at 400Xmagnification submitted to quantitative image analysis are on the right. Antibody stained images are shown in the right side upper rows (labeled 1, 2 & 3), with each 400Xfield analyzed by Nikon Imaging Software in the lower rows (labeled 1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: Runx2 immunostaining in 3-week pellet cultures of primary human OA chondrocytes: WT (GL2 control), IKKα KD cells with empty BIN retroviral vector and IKKα KD cells with kinase-dead murine IKKα(K44M)-BIN. Global views of micromass sphere sections (100Xmagnification) are shown on the left, and three random fields at 400Xmagnification submitted to quantitative image analysis are on the right. Antibody stained images are shown in the right side upper rows (labeled 1, 2 & 3), with each 400Xfield analyzed by Nikon Imaging Software in the lower rows (labeled 1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Retroviral, Plasmid Preparation, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: PLoS ONE

Article Title: IKKα/CHUK Regulates Extracellular Matrix Remodeling Independent of Its Kinase Activity to Facilitate Articular Chondrocyte Differentiation

doi: 10.1371/journal.pone.0073024

Figure Lengend Snippet: COL10 immunostaining in 3-week pellet cultures of primary human OA chondrocytes: WT (GL2 control), IKKα KD cells with empty BIN retroviral vector and IKKα KD cells with kinase-dead murine IKKα(K44M)-BIN. Global views of micromass sphere sections (100Xmagnification) are shown on the left, and three random fields at 400Xmagnification submitted to quantitative image analysis are on the right. Antibody stained images are shown in the right side upper rows (labeled 1, 2 & 3), with each 400Xfield analyzed by Nikon Imaging Software in the lower rows (labeled 1.1, 2.1 & 3.1). For each sample an IHC staining threshold was established based on an antibody isotype control. Antibody staining is in red in conjunction with hematoxylin (blue) nuclear counterstaining. Data are representative examples of multiple sections of micromasses prepared with ACs of 3 OA patients; and results of all such experiments are presented as statistically analyzed bar graphs in .

Article Snippet: Frozen sections (5 μm) of 1-week pellet cultures of human OA chondrocytes were analyzed by immunohistochemistry (IHC) in conjunction with hematoxylin nuclear counterstaining (CAT hematoxylin – BIOCARE Medical) using antibodies against type II collagen (COL2), Runx2, type×collagen (COL10) and the antibody C1,2C (IBEX Inc.) against a specific COL2–3/4C neo-epitope, as previously described .

Techniques: Immunostaining, Control, Retroviral, Plasmid Preparation, Staining, Labeling, Imaging, Software, Immunohistochemistry

Journal: iScience

Article Title: Single-cell transcriptome atlas of male mouse pituitary across postnatal life highlighting its stem cell landscape

doi: 10.1016/j.isci.2024.111708

Figure Lengend Snippet: Pituitary stem cell markers (A) Heatmap displaying scaled expression (see color scale) in the pituitary stem cell clusters compared to all other pituitary cell types (of undamaged pituitary, all ages combined). (B) Immunofluorescence staining of SOX2 (magenta) together with ALDOC or KLF5 (green) in (undamaged) young-adult pituitary and derived organoids. Nuclei are labeled with Hoechst33342 (blue) (scale bar, 100 μm). Boxed area in young-adult pituitary is magnified in right panels. Separate ALDOC and KLF5 signals are shown in organoids in right panels. AL, anterior lobe; PL; posterior lobe. (C) Scatterplot of scaled regulon specificity score (RSS) and scaled mean regulon activity (MRA) in SC1 + SC2 (of undamaged pituitary, all ages combined) with each dot indicating an individual regulon. Specific regulons are indicated by color and annotated genes shown, including AP-1 complex subunits ( Atf family, magenta ; Fos family, cyan; Jun family, orange) and Klf5 (green). (D) UMAP displaying expression of Klf5 target genes, visualized by module score (see color scale). (E) Network visualization of significantly upregulated regulons (rectangles) involved in AP-1 complex and their target genes (ovals) in SC1 + SC2 versus the other cell types ( Atf family, magenta ; Fos family, cyan; Jun family, orange). (F) Organoid development and growth from young-adult AL in the absence (PitOM) or presence of KLF5 inhibitor (KLF5i) ( top ). Organoid development and growth from young-adult AL in the absence (PitOM) or presence of AP-1 inhibitor (AP-1i) ( bottom ). Representative brightfield pictures of organoid cultures on day 10–11 (scale bar, 200 μm; left ) and quantification of organoid area (relative to PitOM at endpoint) over time in culture are shown. Bar plots represent absolute area under the curve (AUC) calculated from organoid area ( n = 3 biological replicates per treatment; right ). Mean ± standard error of the mean (SEM) is depicted. ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001; unpaired, two-tailed Student’s t test. See also Figure S2 , Tables S3 , , and .

Article Snippet: Following incubation with primary and secondary antibodies ( ) and nuclear counterstaining with Hoechst33342 (Merck), sections were mounted with ProLong Gold (Thermo Fisher Scientific, Waltham, MA).

Techniques: Expressing, Immunofluorescence, Staining, Derivative Assay, Labeling, Activity Assay, Two Tailed Test

Journal: iScience

Article Title: Single-cell transcriptome atlas of male mouse pituitary across postnatal life highlighting its stem cell landscape

doi: 10.1016/j.isci.2024.111708

Figure Lengend Snippet: Cellular crosstalk and regulatory circuits in the pituitary stem cell niche (A) Upregulated DEG-associated GO biological processes and terms in SC1 + SC2 versus all other AL cells (undamaged young-adult pituitary). (B) Dotplot displaying predicted ligand-receptor pairs of the EGF family involved in crosstalk between the stem cells (SC1 + SC2, here referred to as SC) and other cell clusters, as inferred by CellPhoneDB (undamaged pituitary, all ages combined). Dot size represents –log10 of p values and color intensity (see scale) indicates mean of average expression of ligand and receptor. (C) Dotplot displaying average expression (color intensity, see scale) of EGF family ligands and receptors and percentage of cells (dot size) within the AL cell clusters expressing the specified gene (undamaged pituitary, all ages combined). (D) Organoid development and growth from young-adult AL in pituitary organoid medium (PitOM; containing EGF), in PitOM without EGF (-EGF), in PitOM without EGF supplemented with NRG1 (-EGF +NRG1) or in PitOM (containing EGF) with NRG1 (+NRG1). Representative brightfield pictures of organoid cultures (day 10; scale bar, 200 μm; left ) are shown. Bar plots represent the calculated AUC of organoid number (count) and total organoid area (biological replicates per treatment, n = 3; right ). Mean ± SEM is depicted. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001; one-way ANOVA with Dunnett’s multiple comparisons test. (E) Dotplot displaying predicted Hgf-Met ligand–receptor interaction between stem cells (SC1 + SC2, here referred to as SC) and corticotropes (Cortico) or mesenchymal cells (MC), as inferred by CellPhoneDB. Dot size represents –log10 of p values and color intensity (see scale) indicates mean of average expression of Hgf and Met . (F) Dotplot displaying average expression (color intensity, see scale) of Hgf and Met and percentage of cells (dot size) within the AL cell clusters expressing Hgf and Met (undamaged pituitary, all ages combined). (G) Organoid number growing from young-adult AL in absence (PitOM) and presence of HGF (+HGF). Representative brightfield pictures of organoid cultures (day 10; scale bar, 200 μm; left ) are shown. Bar plot represents the calculated AUC of organoid number (biological replicates per treatment, n = 6; right ). Mean ± SEM is depicted. ∗∗∗ p ≤ 0.001; unpaired, two-tailed Student’s t test. (H) Organoid number growing from young-adult AL in absence (PitOM) and presence of cMET inhibitor (+cMETi). Representative brightfield pictures of organoid cultures (day 10; scale bar, 200 μm; left ) are shown. Bar plot represents the calculated AUC of organoid number (biological replicates per treatment, n = 6; right ). Mean ± SEM is depicted. ns, non-significant; unpaired, two-tailed Student’s t test. (I) Dotplot displaying predicted Mdk-Ptprz1 interaction between (proliferating) stem cells (SC1 + SC2, here referred to as SC, and Prolif SC) and corticotropes (Cortico), as revealed by CellPhoneDB. Dot size represents –log10 of p values and color intensity (see scale) indicates mean of average expression of Mdk and Ptprz1 . (J) Dotplot displaying average expression (color intensity, see scale) of Mdk and Ptprz1 and percentage of cells (dot size) within the AL cell clusters expressing Mdk and Ptprz1 (undamaged pituitary, all ages combined). (K) Immunofluorescence staining of PTPRZ1 (green; top ) and ACTH (magenta; top ) in young-adult pituitary. Boxed area is magnified. Representative brightfield image of AtT-20 live cell culture ( bottom left ) and immunofluorescence staining of PTPRZ1 in fixed AtT-20 cells (magenta; bottom right ). Nuclei are labeled with Hoechst33342 (blue) (scale bar, 100 μm). AL, anterior lobe. (L) Gene expression level of corticotrope (precursor) markers ( Tbx19 , Pomc) in AtT-20 cells as determined by RT-qPCR in the absence (control (CTRL)) or presence of MDK at increasing concentrations ( n = 3 biological replicates per treatment). Mean ± SEM is depicted. ∗ p ≤ 0.05; two-way ANOVA with Šídák’s multiple comparisons test. See also Figure S3 , Tables S3 , and .

Article Snippet: Following incubation with primary and secondary antibodies ( ) and nuclear counterstaining with Hoechst33342 (Merck), sections were mounted with ProLong Gold (Thermo Fisher Scientific, Waltham, MA).

Techniques: Expressing, Two Tailed Test, Immunofluorescence, Staining, Cell Culture, Labeling, Quantitative RT-PCR, Control

Journal: Progress in retinal and eye research

Article Title: Development of the hyaloid, choroidal and retinal vasculatures in the fetal human eye

doi: 10.1016/j.preteyeres.2017.10.001

Figure Lengend Snippet: VEGFR2 (A–D), CD31 (E–H) and CD34 (I–L) labeling (all red) with epsilon hemoglobin (green, Hb-ε) and DAPI (blue, nuclei) in blood island-like structures in the 6 WG vitreous. Free erythroblasts co-expressed VEGFR2 and Hb-ε (arrowhead in A–D) as did some cells within blood islands (arrow in A–D). All cells of the blood islands expressed VEGFR2. CD31 and Hb-ε were co-expressed in select cells of blood islands (arrow in E–H) while free Hb-ε+ erythroblasts were only weakly immunoreactive for CD31 (arrowhead in E–H). CD34 and Hb-ε were co-expressed in some scattered cells in blood island-like structures (arrows in I–L), while free Hb-ε+ erythroblasts were CD34− (not shown). (Scale bar in A, E & I = 10 μm) (Fig. 5 from McLeod et al., Invest. Ophthalmol. Vis. Sci. 53:7918, 2012, with permission).

Article Snippet: A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) ( from Hasegawa et al., Developmental Dynamics 236:2096 from Hasegawa et al., Developmental Dynamics 236:2007, with permission). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 13 caption a7 CD39 labeled flat choroids shows the pattern of the developing CC in embryonic and fetal human eyes. (A) CD39 + cells are organized into blood island-like structures (circle) at 5.5 WG. (B) Highly cellular linear cord-like structures without apparent lumen have formed at 9 WG (double arrows). (C) The capillaries are narrower, lumens have formed, and cellularity has decreased markedly at 12 WG. (Scale bar = 10 μm) ( from McLeod and Lutty, Chapter 104, page 1506 in The New Visual Neurosciences, Ed.

Techniques: Labeling

Journal: Progress in retinal and eye research

Article Title: Development of the hyaloid, choroidal and retinal vasculatures in the fetal human eye

doi: 10.1016/j.preteyeres.2017.10.001

Figure Lengend Snippet: CD31/Hb-ε double positive cells in developing choroid. (A–D) Clusters of CD31+/Hb-ε+ (red/green) (arrows) were visible in the choriocapillaris layer at 6 WG, whereas in the choroidal stroma had isolated cells (double arrow) that were also double positive. (E–H) There were isolated cells (arrows) that were CD31/Hb-ε positive at 7 WG, which appeared to be attached to the choriocapillaris. Some of these cells were deep in the choroid, resembling those found at 6 WG. Having both CD31/Hb-ε positive in the same cells gives them characteristics of hematopoietic and endothelial cells. B and F are merged images of the single color images C, D and G, H, respectively. A and E are images showing nuclear counter staining with DAPI (blue) merged with B and F, respectively. (Scale bar = 10 μm) (Fig. 4 from Hasegawa et al., Developmental Dynamics 236:2093Fig. 4 from Hasegawa et al., Developmental Dynamics 236:2007, with permission).

Article Snippet: A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) ( from Hasegawa et al., Developmental Dynamics 236:2096 from Hasegawa et al., Developmental Dynamics 236:2007, with permission). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 13 caption a7 CD39 labeled flat choroids shows the pattern of the developing CC in embryonic and fetal human eyes. (A) CD39 + cells are organized into blood island-like structures (circle) at 5.5 WG. (B) Highly cellular linear cord-like structures without apparent lumen have formed at 9 WG (double arrows). (C) The capillaries are narrower, lumens have formed, and cellularity has decreased markedly at 12 WG. (Scale bar = 10 μm) ( from McLeod and Lutty, Chapter 104, page 1506 in The New Visual Neurosciences, Ed.

Techniques: Isolation, Staining

Journal: Progress in retinal and eye research

Article Title: Development of the hyaloid, choroidal and retinal vasculatures in the fetal human eye

doi: 10.1016/j.preteyeres.2017.10.001

Figure Lengend Snippet: CD34/Ki67 staining of fetal choroid. There are no CD34 (red) and Ki67 (green) double stained cells at 6 WG (A, B), while CD34/Ki67 double positive cells (arrow) were observed in a forming intermediate vessel at 12 WG (C, D). B and D are merged images of CD34 (Red) and Ki67 (Green). A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) (Fig. 8 from Hasegawa et al., Developmental Dynamics 236:2096Fig. 8 from Hasegawa et al., Developmental Dynamics 236:2007, with permission).

Article Snippet: A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) ( from Hasegawa et al., Developmental Dynamics 236:2096 from Hasegawa et al., Developmental Dynamics 236:2007, with permission). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 13 caption a7 CD39 labeled flat choroids shows the pattern of the developing CC in embryonic and fetal human eyes. (A) CD39 + cells are organized into blood island-like structures (circle) at 5.5 WG. (B) Highly cellular linear cord-like structures without apparent lumen have formed at 9 WG (double arrows). (C) The capillaries are narrower, lumens have formed, and cellularity has decreased markedly at 12 WG. (Scale bar = 10 μm) ( from McLeod and Lutty, Chapter 104, page 1506 in The New Visual Neurosciences, Ed.

Techniques: Staining

Journal: Progress in retinal and eye research

Article Title: Development of the hyaloid, choroidal and retinal vasculatures in the fetal human eye

doi: 10.1016/j.preteyeres.2017.10.001

Figure Lengend Snippet: Cryosection of a 7 WG eye immunolabeled with anti-CD39 (red) and anti-CXCR4 (green) and counterstained with DAPI (blue). The fetal vasculature in vitreous and the CC prominently express CD39 as well as angioblasts in inner retina. The inner neuroblastic layer cells, as well as some cells in inner retina, express CXCR4. Scale bar = 50 μm.

Article Snippet: A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) ( from Hasegawa et al., Developmental Dynamics 236:2096 from Hasegawa et al., Developmental Dynamics 236:2007, with permission). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 13 caption a7 CD39 labeled flat choroids shows the pattern of the developing CC in embryonic and fetal human eyes. (A) CD39 + cells are organized into blood island-like structures (circle) at 5.5 WG. (B) Highly cellular linear cord-like structures without apparent lumen have formed at 9 WG (double arrows). (C) The capillaries are narrower, lumens have formed, and cellularity has decreased markedly at 12 WG. (Scale bar = 10 μm) ( from McLeod and Lutty, Chapter 104, page 1506 in The New Visual Neurosciences, Ed.

Techniques: Immunolabeling

Journal: Progress in retinal and eye research

Article Title: Development of the hyaloid, choroidal and retinal vasculatures in the fetal human eye

doi: 10.1016/j.preteyeres.2017.10.001

Figure Lengend Snippet: Cryosections (A–D) and a whole mount retina (E–F) from 7 WG eyes. (A) Near the nerve head (NH), fusiform-shaped cells in the nerve fiber layer (NFL) were positive for both CXCR4 (green) and CD39 (red) (arrow in A–B). Inset shows the double-labeled cell indicated by the arrow in “A”–“B” at higher magnification. (DAPI counterstain showing the blue nuclei, red CD39 and green CXCR4 channels) (B) Same section without the DAPI channel shows CXCR4 immunostained cells the entire inner neuroblastic layer and some double-labeled cells in the nerve fiber layer express CXCR4. A few double-labeled cells are near the inner portion of the INL. (C) Same area in a serial section labeled for CXCR4 (green) and Ki67 (red). Only the outer neuroblastic layer is Ki67+. (D) Same section with the CXCR4 green channel turned off. (E) Retinal whole mount shows fusiform-shaped cells (arrows) near the optic nerve head (to the right and out of the field) that express both CD39 (red) and CXCR4 (green). (F) Same region as “E” with only the green CXCR4 channel shown. (A–D scale bar = 20 μm; E–F scale bar = 40 μm).

Article Snippet: A and C are merged B and D with DAPI nuclear counterstaining (blue). (Bar = 10 μm) ( from Hasegawa et al., Developmental Dynamics 236:2096 from Hasegawa et al., Developmental Dynamics 236:2007, with permission). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 13 caption a7 CD39 labeled flat choroids shows the pattern of the developing CC in embryonic and fetal human eyes. (A) CD39 + cells are organized into blood island-like structures (circle) at 5.5 WG. (B) Highly cellular linear cord-like structures without apparent lumen have formed at 9 WG (double arrows). (C) The capillaries are narrower, lumens have formed, and cellularity has decreased markedly at 12 WG. (Scale bar = 10 μm) ( from McLeod and Lutty, Chapter 104, page 1506 in The New Visual Neurosciences, Ed.

Techniques: Labeling