Journal: Development (Cambridge, England)

Article Title: Newborn horizontal cells migrate bi-directionally across the neuroepithelium during retinal development.

doi: 10.1242/dev.01018

Figure Lengend Snippet: Fig. 1. Fluorescence immunohistochemistry for Lim1 and Prox1 in stage 24-35 developing avian retina. Micrographs showing immunohistochemistry for (A- H) Lim1 (green) in horizontal cells (HCs) and Ng-CAM (red) on ganglion cells in stage 24-35 retina. (I) Overview of Lim1 immunoreactivity in stage 35 retina. The gradual spatial maturation with corresponding developmental stages are indicated. (J-M) Prox1 (red) and Lim1 (green) immunoreactivity in (J, K) stage 28, (L) stage 33 and (M) stage 35 retinas. Arrows in I and J indicate the central region of the retina. st, stage. Scale bars: in A (for A-C), D (for D-H), K, L (for L-M) are 20 µm and I (for I-J) is 100 µm.

Article Snippet: Primary antibodies used in this study were directed towards Lim1/2 (1:5-50, 4F2, Developmental Studies Hybridoma Bank DSHB), Ng-CAM [1:2000, (de la Rosa et al., 1990)], Prox1 (1:4000, gift from Dr M. Nakafuku), GABA (1:1000, A2052; 1:500, A0310, Sigma), Pax6 (1:200, PAX6, DSHB), Lim3 (1:200, 67.4E12, DSHB), Ap2α (1:200, 3B5, DSHB), Islet1 (1:200, 40.2D6, DSHB), Chx10 (1:4000, gift from Dr J. Ericson), calretinin (1:1000, 1741-1007, Anawa, Zürich), Brn3a (1:100, MAB1585, Chemicon), Brn3b (1:200, sc-6026, Santa Cruz) and BrdU (1:50, M20105S, Biosite).

Techniques: Fluorescence, Immunohistochemistry

Journal: Development (Cambridge, England)

Article Title: Newborn horizontal cells migrate bi-directionally across the neuroepithelium during retinal development.

doi: 10.1242/dev.01018

Figure Lengend Snippet: Fig. 3. GFP and Lim1 immunohistochemistry in biolistically transfected stage 31 retinas. (A-C) A time-lapse series of a retinal slice from a biolistically transfected stage 31 retina using a green fluorescent protein (GFP) expression vector. A cell indicated by a large arrowhead is migrating towards the ventricular side in relation to its starting point (small arrowhead). (D) Lim1 (red) and GFP (green) co-labelling in cells fixed and cryosectioned 24 hours after gene transfer. (E-H) GFP (green), and other markers (red) in fixed and cryosectioned retinas at different time points after transfection. White arrows indicate gold particles in the internal retina. Black arrows indicate GFP and antibody co-expression. (E) GFP and Prox1 22 hours after transfection. (F) Ganglion cell marker Ng-CAM (G4) expression and gold particle penetration 15 minutes after transfection. GFP is not yet expressed. (G) Ganglion cell marker Islet1 and GFP, 22 hours after transfection. (H) GFP and Ng-CAM (G4) in a retina 18 hours after transfection. Gold particles as well as cells expressing GFP are found on the ventricular side of the retina (white arrow). Scale bars: in A (for A-C), D,E (for E-H) 20 µm.

Article Snippet: Primary antibodies used in this study were directed towards Lim1/2 (1:5-50, 4F2, Developmental Studies Hybridoma Bank DSHB), Ng-CAM [1:2000, (de la Rosa et al., 1990)], Prox1 (1:4000, gift from Dr M. Nakafuku), GABA (1:1000, A2052; 1:500, A0310, Sigma), Pax6 (1:200, PAX6, DSHB), Lim3 (1:200, 67.4E12, DSHB), Ap2α (1:200, 3B5, DSHB), Islet1 (1:200, 40.2D6, DSHB), Chx10 (1:4000, gift from Dr J. Ericson), calretinin (1:1000, 1741-1007, Anawa, Zürich), Brn3a (1:100, MAB1585, Chemicon), Brn3b (1:200, sc-6026, Santa Cruz) and BrdU (1:50, M20105S, Biosite).

Techniques: Immunohistochemistry, Transfection, Expressing, Plasmid Preparation, Marker

Journal: Development (Cambridge, England)

Article Title: Newborn horizontal cells migrate bi-directionally across the neuroepithelium during retinal development.

doi: 10.1242/dev.01018

Figure Lengend Snippet: Fig. 5. Lamination of the stage 35 retina as shown by transcription factor immunoreactivity. Representative fluorescence micrographs showing transcription factor immunoreactivity in the central region of stage 35 retina. (A) Pax6, (B) Prox1, (D) Ap2α, (E) Prox1, (G) Lim3, (H) Prox1, (J) Lim3, (K) Chx10, (M) Lim1 and (N) Chx10. (C,F,I,L,O) Merged image of the fluorescence micrographs presented in the top two rows. (P) Schematic summary of the immunoreactivity shown in A-O. (+) Moderate and (++) strong immunoreactivity. The results are representative of and were collected from more than three animals in each case. ONL/onl: outer nuclear layer, INL/inl: inner nuclear layer (a: outer external INL, b: inner external INL, c: outer internal INL, d: inner internal INL), GCL/glc: ganglion cell layer. Scale bar: A (for A-O) 20 µm.

Article Snippet: Primary antibodies used in this study were directed towards Lim1/2 (1:5-50, 4F2, Developmental Studies Hybridoma Bank DSHB), Ng-CAM [1:2000, (de la Rosa et al., 1990)], Prox1 (1:4000, gift from Dr M. Nakafuku), GABA (1:1000, A2052; 1:500, A0310, Sigma), Pax6 (1:200, PAX6, DSHB), Lim3 (1:200, 67.4E12, DSHB), Ap2α (1:200, 3B5, DSHB), Islet1 (1:200, 40.2D6, DSHB), Chx10 (1:4000, gift from Dr J. Ericson), calretinin (1:1000, 1741-1007, Anawa, Zürich), Brn3a (1:100, MAB1585, Chemicon), Brn3b (1:200, sc-6026, Santa Cruz) and BrdU (1:50, M20105S, Biosite).

Techniques:

Journal: PLoS ONE

Article Title: Ctip2-, Satb2-, Prox1-, and GAD65-Expressing Neurons in Rat Cultures: Preponderance of Single- and Double-Positive Cells, and Cell Type-Specific Expression of Neuron-Specific Gene Family Members, Nsg-1 (NEEP21) and Nsg-2 (P19)

doi: 10.1371/journal.pone.0140010

Figure Lengend Snippet: Commercially available primary antibodies.

Article Snippet: Mouse anti-PROX1 , Active Motif , 61093 , 1 to 1000.

Techniques:

Journal: eLife

Article Title: Fish primary embryonic pluripotent cells assemble into retinal tissue mirroring in vivo early eye development

doi: 10.7554/eLife.66998

Figure Lengend Snippet: ( a ) Scheme of organoid generation from Atoh7::EGFP transgenic fish. ( b ) Fluorescent images of day 3 Atoh7::EGFP organoids (generated by aggregation of <1000 cells). ( c ) Optical sections and maximal projections showing EGFP expression in the eye of the developing embryo at 2 dpf and the retinal organoid at day 3 co-stained with antibody against HuC/D and DAPI. ( d ) Optical sections showing expression of HuC/D (amacrine and ganglion cells), Otx2 (bipolar and photoreceptor cells), and Prox1 (horizontal cells) in day 4 organoid. ( e ) Sketch of the arrangement of cellular layers in the organoid and the embryonic retina. dpf, days post-fertilization. Scale bar: 100 μm.

Article Snippet: Antibody , Anti-Prox1 (polyclonal rabbit IgG) , Merck , Cat#:AB5475 RRID: AB_177485 , IHC (1:500).

Techniques: Transgenic Assay, Generated, Expressing, Staining

Journal: eLife

Article Title: Fish primary embryonic pluripotent cells assemble into retinal tissue mirroring in vivo early eye development

doi: 10.7554/eLife.66998

Figure Lengend Snippet: ( a ) Scheme of organoid generation from Atoh7::EGFP transgenic fish. ( b ) Bright-field and fluorescent images of day 3 Atoh7::EGFP organoids. ( c ) Optical sections of day 4 Atoh7::EGFP organoids co-stained with antibodies against HuC/D (amacrine and ganglion cells), Otx2 (bipolar and photoreceptor cells), and Prox1 (horizontal cells). Dashed lines indicate the retinal domain. Scale bar: 100 μm.

Article Snippet: Antibody , Anti-Prox1 (polyclonal rabbit IgG) , Merck , Cat#:AB5475 RRID: AB_177485 , IHC (1:500).

Techniques: Transgenic Assay, Staining

Journal: eLife

Article Title: Fish primary embryonic pluripotent cells assemble into retinal tissue mirroring in vivo early eye development

doi: 10.7554/eLife.66998

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-Prox1 (polyclonal rabbit IgG) , Merck , Cat#:AB5475 RRID: AB_177485 , IHC (1:500).

Techniques: Recombinant, Plasmid Preparation, Sequencing, Software

Journal: Cancers

Article Title: CD147 Promotes Tumor Lymphangiogenesis in Melanoma via PROX-1

doi: 10.3390/cancers13194859

Figure Lengend Snippet: CD147 regulates LECs lymphangiogenic markers at transcriptional and translational levels. ( a ) qRT-PCR of FLT4 , PROX1 , LYVE1 , and PDPN expression in LECs treated or not with rhCD147, using PPIA as a reference. Columns indicate the means of three independent experiments carried out in triplicate; bars indicate SD; * p < 0.05, ** p < 0.005, *** p < 0.0001. ( b ) Western blot analyses of VEGFR-3, PROX-1, LYVE-1, Podoplanin, and CD147 expression in total lysates of LECs treated or not with rhCD147. ( c ) Three independent Western blots were quantified for densitometry band analyses. Equal loading of proteins was assessed by probing for β-actin. Bars, SD; * p < 0.05, ** p <0.005, *** p < 0.0001. ( d ) Representative confocal images of PROX-1, LYVE-1 and Podoplanin immunofluorescent staining on LECs treated or not with rhCD147 (63X magnification). Scale bar: 30 µm. ( e ) ChIP-qPCR was performed on LECs treated or not with rhCD147, with antibody against PROX-1 total and control IgG. Columns indicate means of two independent experiments; and bars, SD; ns, non significant, * p < 0.05, ** p < 0.005, *** p < 0.0001.

Article Snippet: Then, 20 µg of lysate in 4× Laemmli per lane were resolved by reducing 10% SDS-PAGE followed by Western blot analyses with either Rabbit anti-LYVE-1 polyclonal antibody (#ab14917, Abcam, Cambridge, UK), Rabbit anti-VEGFR-3 polyclonal antibody (#102-PA22AG, ReliaTech), Rabbit anti-PROX-1 polyclonal antibody (#07-537, Merck), Mouse anti-Podoplanin monoclonal antibody (#ab10288, Abcam, Cambridge, UK) or Mouse monoclonal anti-β-actin antibody (#A2228, Merck) for loading control.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining

Journal: Cancers

Article Title: CD147 Promotes Tumor Lymphangiogenesis in Melanoma via PROX-1

doi: 10.3390/cancers13194859

Figure Lengend Snippet: CD147 regulates LECs lymphangiogenic markers through PROX-1 transcription factor. LECs were transfected with scrambled or PROX-1 siRNA and treated or not with rhCD147. ( a ) qRT-PCR of PROX1 , PDPN , and LYVE1 expression using PPIA as a reference. Columns indicate means of three independent experiments carried out in triplicate; bars indicate SD; * p < 0.05, *** p < 0.0001 for siRNA-PROX-1 vs. siRNA-scrambled. ns, nonsignificant, # p < 0.05, ### p < 0.0001 for treated vs. untreated cells. ( b ) Representative confocal images of immunofluorescence staining for PROX-1, Podoplanin and LYVE-1 (63× magnification).

Article Snippet: Then, 20 µg of lysate in 4× Laemmli per lane were resolved by reducing 10% SDS-PAGE followed by Western blot analyses with either Rabbit anti-LYVE-1 polyclonal antibody (#ab14917, Abcam, Cambridge, UK), Rabbit anti-VEGFR-3 polyclonal antibody (#102-PA22AG, ReliaTech), Rabbit anti-PROX-1 polyclonal antibody (#07-537, Merck), Mouse anti-Podoplanin monoclonal antibody (#ab10288, Abcam, Cambridge, UK) or Mouse monoclonal anti-β-actin antibody (#A2228, Merck) for loading control.

Techniques: Transfection, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Early Seizure Activity Accelerates Depletion of Hippocampal Neural Stem Cells and Impairs Spatial Discrimination in an Alzheimer’s Disease Model

doi: 10.1016/j.celrep.2019.05.101

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-Prox1 , PhosphoSolutions , Cat#: 1685-Prox1; RRID: AB_2492217.

Techniques: Recombinant, Imaging, Software, Microscopy