goat anti prox1  (R&D Systems)

Journal: Blood

Article Title: Aberrant mural cell recruitment to lymphatic vessels and impaired lymphatic drainage in a murine model of pulmonary fibrosis

doi: 10.1182/blood-2011-12-396895

Figure Lengend Snippet: (A) Immunodetection of mural cells (SMA) and Lyve-1–positive vessels in fibrotic lungs of mice treated with bleomycin (Bleo; 10 mg/kg body weight twice a week intraperitoneally) and in healthy controls of mice treated with PBS (100 μL twice a week intraperitoneally) at day 28. Fibrosis is shown at early, intermediate, and late stage. Scale bars equal 100 μm. (B) Quantitative analysis of Lyve-1–positive vessels with SMA-positive mural cells in lung tissue of bleomycin-injected and healthy control mice at day 28 (controls n = 6; Bleo n = 8). (C) Quantitative analysis of VEGFR3-positive lymphatic vessels with SMA-positive mural cells in lung tissue of bleomycin-injected and healthy control mice at day 28 (controls n = 5; Bleo n = 5). (D) Quantitative analysis of Prox1-positive lymphatic vessels with SMA-positive mural cells in lung tissue of bleomycin-injected and healthy control mice at day 28 (controls n = 5; Bleo n = 5). Error bars show SEM.

Article Snippet: Primary antibodies used in this study were as follows: rabbit polyclonal to Lyve-1 at a 1:200 dilution (Abcam), anti-laminin antibody at a 1:50 dilution (Sigma-Aldrich), goat polyclonal to mouse serum albumin at a 1:500 dilution (Abcam), mouse mAb phospho-PDGFR-β at a 1:100 dilution (Cell Signaling), rabbit monoclonal antibody (mAb) PDGFR-β at a 1:100 dilution (Cell Signaling), rabbit polyclonal to PDGF BB at a 1:100 dilution (Abcam), rat anti–mouse VEGFR3 at a 1:100 dilution (BD Bioscience), human Prox1 affinity purified polyclonal AB goat immunoglobulin (Ig) G at a 1:200 dilution (R&D Systems), anti–mouse α-SMA at a 1:500 dilution (Chemicon), biotinylated hyaluronan-binding peptide at a 1:100 dilution (Calbiochem), D2-40 and S100A4 (FSP) at a 1:200 dilution (Dako).

Techniques: Immunodetection, Injection, Control

Journal: Blood

Article Title: Aberrant mural cell recruitment to lymphatic vessels and impaired lymphatic drainage in a murine model of pulmonary fibrosis

doi: 10.1182/blood-2011-12-396895

Figure Lengend Snippet: (A) Quantitative analysis of the PDGFR-P expression in isolated lymphatic endothelial cells of bleomycin-treated (Bleo) and healthy control mice at day 28 (controls n = 15; Bleo n = 14). Immunostaining of PDGF-B (B) or PDGFR-β (C) and lymphatic vessels (VEGFR3) on lung tissue of fibrotic and control mice at day 28. Scale bars equal 100 μm. (D) Westernblot of phospho–PDGFR-β and total PDGFR-β of whole lung lysates at day 28 after treatment with bleomycin or PBS, respectively (controls n = 5; Bleo n = 13). (E) Quantitative analysis of the ratio of phospho–PDGFR-β to total PDGFR-β at day 28 after treatment with bleomycin or PBS (controls n = 5, Bleo n = 13). (F) Double-immunodetection of phospho–PDGFR-β and lymphatic vessels (VEGFR3) on healthy and fibrotic murine lung tissue at day 28. Scale bars equal 100 μm. Quantitative analysis of Lyve-1–positive lymphatic vessels (G), VEGFR3-positive lymphatic vessels (H) or Prox1-positive lymphatic vessels (I) covered with SMA-positive mural cells at day 28 after treatment with bleomycin, bleomycin + AG-1296 (Bleo + AG-1296; 10 mg/kg body weight twice a week intraperitoneally; 0.5 ng, 5 times per week intraperitoneally, starting at day 14 after first bleomycin injection) or in healthy control lungs of PBS-treated mice (controls n = 15; Bleo n = 29, Bleo + AG-1296, n = 13). Error bars show SEM.

Article Snippet: Primary antibodies used in this study were as follows: rabbit polyclonal to Lyve-1 at a 1:200 dilution (Abcam), anti-laminin antibody at a 1:50 dilution (Sigma-Aldrich), goat polyclonal to mouse serum albumin at a 1:500 dilution (Abcam), mouse mAb phospho-PDGFR-β at a 1:100 dilution (Cell Signaling), rabbit monoclonal antibody (mAb) PDGFR-β at a 1:100 dilution (Cell Signaling), rabbit polyclonal to PDGF BB at a 1:100 dilution (Abcam), rat anti–mouse VEGFR3 at a 1:100 dilution (BD Bioscience), human Prox1 affinity purified polyclonal AB goat immunoglobulin (Ig) G at a 1:200 dilution (R&D Systems), anti–mouse α-SMA at a 1:500 dilution (Chemicon), biotinylated hyaluronan-binding peptide at a 1:100 dilution (Calbiochem), D2-40 and S100A4 (FSP) at a 1:200 dilution (Dako).

Techniques: Expressing, Isolation, Control, Immunostaining, Immunodetection, Injection

Journal: PLoS ONE

Article Title: Comprehensive Expression of Wnt Signaling Pathway Genes during Development and Maturation of the Mouse Cochlea

doi: 10.1371/journal.pone.0148339

Figure Lengend Snippet: (A) Transverse section through the cochlea on E12.5. Sfrp3 was expressed in the prosensory epithelia of the cochlea. (B) Transverse section through the basal turn on E15.5, Sfrp3 expression was detected in the LER. (C) Transverse section through the basal turn on E17.5, Sfrp3 expression was localized to the LER, outer hair cells and Deiters’ cells. (D) Transverse section through the basal turn on P0. Sfrp3 expression was maintained in the outer hair cells and Deiters’ cells. Inset image shows a high magnification view the image in D, counterstained by immunofluorescence for hair cell marker Myo7a (red) and supporting cell marker, Prox1 (green), Sfrp3 expression is detected in the outer pillar cell, the first and second Deiter’s cells but not in the inner pillar or third Deiters’ cell. (E) Transverse section through apical turn on P4. Sfrp3 was expressed in the outer hair cells and Claudius’ cells. (F) Transverse section through the basal turn on P6. Sfrp3 expression was restricted to the inner sulcus and Claudius’ cells. (G) A transverse section through a P30 cochlea. Sfrp3 expression was maintained in Claudius cells and the inner sulcus. (H) A transverse section through the apical turn of the cochlea on P0. Wnt5a mRNA expression localizes to the GER. (I) Transverse section adjacent to section used in (H). Sfrp3 expression was detected at high levels in the sensory epithelium and the LER. (J) Image (H) overlaid with Image (I), demonstrating that Sfrp3 and Wnt5a are reciprocally expressed. Hair cells were labeled by Myo7a (red). Abbreviations: A: anterior; P: posterior; D: dorsal; V: ventral; PSE: prosensory epithelium; GER: greater epithelial ridge; LER: lesser epithelial ridge; inner hair cell (shown as asterisk *); IP: inner pillar cell; OP: outer pillar cell; OHC: outer hair cell; Ds: Deiters’ cells; H: Hensen’s cells; C: Claudius’ cells; SV: stria vascularis; and RM: Reissner’s membrane.

Article Snippet: Primary antibody: Rabbit Myosin-7a (Myo7a) (1:1000; Proteus Biosciences) and Goat Prox1 (1:500; R&D Systems).

Techniques: Expressing, Immunofluorescence, Marker, Labeling, Membrane