Journal: BMC Neuroscience

Article Title: Chronic exposure to high fat diet triggers myelin disruption and interleukin-33 upregulation in hypothalamus

doi: 10.1186/s12868-019-0516-6

Figure Lengend Snippet: List of antibodies in the study for immunofluorescence or western blot analysis

Article Snippet: Polyclonal goat anti-IL-33 , R and D Systems Cat#AF3626, RRID:AB_884269 , Escherichia coli -derived recombinant mouse IL-33 , 1:200 (IF).

Techniques: Immunofluorescence, Western Blot, Purification, Affinity Chromatography, Recombinant, Derivative Assay

Journal: Cell Proliferation

Article Title: Proliferation of myofibroblasts in the stroma of renal oncocytoma

doi: 10.1111/j.1365-2184.2010.00681.x

Figure Lengend Snippet: Protocols for immunostaining

Article Snippet: To reduce non‐specific background staining, sections were pre‐incubated with either (i) normal rabbit (X0902; Dako, Glostrup, Denmark) or swine serum (X0901; Dako, Glostrup, Denmark) at dilution of 1:25 for 15 min, or (ii) 3% bovine serum albumin (Sigma, St Louis, MO, USA) at dilution of 1:25 for 15 min. After washing in PBS, sections were incubated for 35 min with primary antibodies against: (i) MIB‐1 (M7240; Dako, Glostrup, Denmark) at 1:100 dilution, (ii) active caspase 3 (AF835; R&D, Minneapolis, MN, USA) at 1:100 dilution, (iii) SMA (A2547; Sigma) 1:2000 dilution, (iv) β‐catenin (610154; BD, San Jose, CA, USA) 1:100 dilution, (v) E‐cadherin (SC8426; Santa‐Cruz, CA, USA) 1:30 dilution, (vi) collagen I (1310‐01; Southern Biotech, Birmingham, AL, USA) 1:300 dilution, (vii) collagen III (ab6310; ABcam, Cambridge, UK) 1:600 dilution, (viii) fibronectin (ab2413; ABcam, Cambridge, UK) 1:300 dilution, (ix) desmin (M0760; Dako, Glostrup, Denmark) 1:50 dilution and (x) human caldesmon (M3557; Dako, Glostrup, Denmark) 1:200 dilution.

Techniques: Plasmid Preparation

Journal: Cell Proliferation

Article Title: Proliferation of myofibroblasts in the stroma of renal oncocytoma

doi: 10.1111/j.1365-2184.2010.00681.x

Figure Lengend Snippet: Analysis of tumour stroma of oncocytomas of the kidney. Renal oncocytomas were composed of two independent compartments, benign oncocytes and a pronounced fibrotic stroma (desmoplasia) and were associated with excessive deposition of extracellular matrix, which was periodic acid Schiff‐positive (a, pink, asterisk), collagen I‐positive (b, brown, asterisk), fibronectin‐positive (c, brown, asterisk) and collagen III‐positive (d, brown, asterisk), but negative for desmin (e) and human caldesmon (g) immunostaining. Archival smooth muscle tissue was used as positive control for desmin (f, brown, asterisk) and human caldesmon immunostaining (h, brown, asterisk) (magnification, ×400).

Article Snippet: To reduce non‐specific background staining, sections were pre‐incubated with either (i) normal rabbit (X0902; Dako, Glostrup, Denmark) or swine serum (X0901; Dako, Glostrup, Denmark) at dilution of 1:25 for 15 min, or (ii) 3% bovine serum albumin (Sigma, St Louis, MO, USA) at dilution of 1:25 for 15 min. After washing in PBS, sections were incubated for 35 min with primary antibodies against: (i) MIB‐1 (M7240; Dako, Glostrup, Denmark) at 1:100 dilution, (ii) active caspase 3 (AF835; R&D, Minneapolis, MN, USA) at 1:100 dilution, (iii) SMA (A2547; Sigma) 1:2000 dilution, (iv) β‐catenin (610154; BD, San Jose, CA, USA) 1:100 dilution, (v) E‐cadherin (SC8426; Santa‐Cruz, CA, USA) 1:30 dilution, (vi) collagen I (1310‐01; Southern Biotech, Birmingham, AL, USA) 1:300 dilution, (vii) collagen III (ab6310; ABcam, Cambridge, UK) 1:600 dilution, (viii) fibronectin (ab2413; ABcam, Cambridge, UK) 1:300 dilution, (ix) desmin (M0760; Dako, Glostrup, Denmark) 1:50 dilution and (x) human caldesmon (M3557; Dako, Glostrup, Denmark) 1:200 dilution.

Techniques: Immunostaining, Positive Control

Journal: Arthritis Research & Therapy

Article Title: Connective tissue growth factor promotes articular damage by increased osteoclastogenesis in patients with rheumatoid arthritis

doi: 10.1186/ar2863

Figure Lengend Snippet: Connective tissue growth factor expression in synovial tissue of patients with rheumatoid arthritis. The representative results of HE staining (Figure 2A), immunofluorescence anti-CTGF antibody staining (Figure 2B; green), and anti-F4/80 antibody staining (Figure 2C; red) are shown using surgical samples from RA and OA patients. The samples were counterstained by DAPI (blue) for nuclear staining and merge images are shown (Figure 2D). A strong expression of CTGF and F4/80 was observed in the samples of RA compared to OA and the CTGF expression cells were not overlapped with F/40 expression cells indicating that CTGF is upregulated in synovial fibroblasts rather than macrophages.

Article Snippet: The serum level of CTGF in human sera was evaluated by a sandwich ELISA system using two different anti-human CTGF antibodies; monoclonal anti-human CTGF antibody (R&D System, Cat#MAB660) and biotinated anti-human CTGF antibody (R&D System, Cat#BAF660).

Techniques: Expressing, Staining, Immunofluorescence

Journal: Arthritis Research & Therapy

Article Title: Connective tissue growth factor promotes articular damage by increased osteoclastogenesis in patients with rheumatoid arthritis

doi: 10.1186/ar2863

Figure Lengend Snippet: Synergistic effects of connective tissue growth factor on M-CSF/sRANKL-mediated osteoclastic function. Figure 5A shows the results of the resorption of osteoclasts on calcium phosphate. Vacant regions indicated by arrows represent the areas where the osteoclasts actually absorbed. There was no vacant region in negative control cells (M-CSF alone). In contrast to negative control, significant vacant regions were observed in osteoclasts induced by M-CSF/sRANKL. CTGF further expanded the vacant areas in combination with M-CSF/sRANKL and anti-CTGF antibody neutralized this effect. Figure 5B shows the levels of expression of osteoclasts specific proteases (MMP-9 and cathepsin-K) measured by quantitative real time RT-PCR. Synergistic effect of CTGF was also observed for M-CSF/sRANKL-mediated osteoclastogenesis. Bars in Figure 5B indicate the SD.

Article Snippet: The serum level of CTGF in human sera was evaluated by a sandwich ELISA system using two different anti-human CTGF antibodies; monoclonal anti-human CTGF antibody (R&D System, Cat#MAB660) and biotinated anti-human CTGF antibody (R&D System, Cat#BAF660).

Techniques: Negative Control, Expressing, Quantitative RT-PCR

Journal: Arthritis Research & Therapy

Article Title: Connective tissue growth factor promotes articular damage by increased osteoclastogenesis in patients with rheumatoid arthritis

doi: 10.1186/ar2863

Figure Lengend Snippet: Connective tissue growth factor mediate ERK1/2 and focal adhesion kinase activation through integrin αVβ3 signal transduction. Figure 6A shows the immnoprecipitation and immunoblotting analysis. The cell extracts of osteoclasts stimulated with recombinant CTGF (10 or 50 ng/ml) at 60 minutes were precipitated using anti-integrin αVβ3 antibody and subsequently blotted with anti- phosphorylated ERK1/ERK2, conventional ERK1/ERK2, and integrin αVβ3 antibodies respectively. The phosphorylated ERK1/ERK2 was recruited with integrin αVβ3 by CTGF stimulation. Figure 6B shows the immunoblotting analysis using anti- phosphorylated FAK, conventional FAK, and β-actin antibodies in the osteoclasts extracts treated with CTGF (10 ng/ml) at 5, 15, 60, and 120 minutes in the presence or absence of anti-CTGF antibody (1 μg/ml). CTGF stimulation resulted in phosphorylation of FAK from 60 minutes and this effect was neutralized by anti-CTGF antibody suggesting activation of signal transduction pathways through integrin αVβ3.

Article Snippet: The serum level of CTGF in human sera was evaluated by a sandwich ELISA system using two different anti-human CTGF antibodies; monoclonal anti-human CTGF antibody (R&D System, Cat#MAB660) and biotinated anti-human CTGF antibody (R&D System, Cat#BAF660).

Techniques: Activation Assay, Transduction, Western Blot, Recombinant, Phospho-proteomics

Journal: Developmental Cell

Article Title: Lumen Expansion Facilitates Epiblast-Primitive Endoderm Fate Specification during Mouse Blastocyst Formation

doi: 10.1016/j.devcel.2019.10.011

Figure Lengend Snippet: EPI and PrE Expression Levels Are Reduced in ATP1-Inhibited Embryos (A) Immunofluorescence images of TE (Cdx2), EPI (Sox2), and PrE (Gata4) fate in pre-treatment control (E3.5 WT), Atp1 inhibited (E4.0 500 μM and E4.0 250 μM), and end-stage control (E4.0 DMSO) embryos. Lumen boundaries outlined by dashed white line and mean lumen volume in white text. Scale bars, 10 μm. (B) Boxplot of lumen volume for E3.5 WT (N = 21), E4.0 DMSO (N = 24), E4.0 250 μM Atp1 inhibited (N = 14) and E4.0 500 μM Atp1 inhibited (N = 31) embryos indicating that the impact on lumen volume is concentration dependent. (C) Boxplot of fluorescence levels of Cdx2 (gray), Sox2 (green), and Gata4 (magenta) in E4.0 500 μM Atp1 inhibited embryos compared to E4.0 DMSO controls. (D) Schematic 2D representation of 3D analysis method for spatial segregation of ICM lineages. P 1,2,3,4 are 3D points. L ↔ is a 3D line ( P 1 P 2 ↔ equivalent) that defines the embryonic-abembryonic axis. d ‾ is the 3D line segment ( P 3 P 4 ‾ equivalent) that measures the perpendicular distance from the center of a cell to L ↔ . See Image Analysis for formal definitions of all geometric entities. (E) Boxplot of spatial overlap between EPI and PrE lineages within E4.0 control (DMSO, N = 15), E4.0 Atp1 inhibited (500 μM, N =13) and simulated data of maximal overlap in E4.0 WT embryos (Simulation, N = 27). ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05. n.s., not significant. For boxplots: central mark indicates the median; lower edge, 25%; upper edge, 75%; lower whisker, Q1 − (1.5 × IQR), where IQR = Q3 − Q1; upper whisker, Q3 + (1.5 × IQR). See also and ; .

Article Snippet: The following primary antibodies were used in this study: rabbit anti-pERM (Cell Signaling, 3726), mouse anti-Cdx2 (BioGenex, MU392A-UC), goat anti-Sox2 (Santa Cruz Biotechnology, sc-17320), goat anti-Sox2 (R&D Systems, AF2018-SP), rabbit anti-Sox2 (Cell Signaling, 23064), rabbit anti Gata4 (Santa Cruz Biotechnology, sc-9053), goat anti-Gata4 (R&D Systems, AF2606-SP), rabbit anti-GFP (MBL, 598), mouse anti-Rab11 (BD Biosciences; 610656), rat anti-integrin-β1 (Merck Millipore, MAB1997), goat anti-biotinylated Gata4 (R&D Systems, BAF2606) mouse anti-Oct3/4 (Santa Cruz Biotechnology, sc-5279) and mouse anti-Hsp47 (Enzo Life Sciences, M16.10A1).

Techniques: Expressing, Immunofluorescence, Control, Concentration Assay, Fluorescence, Whisker Assay

Journal: Developmental Cell

Article Title: Lumen Expansion Facilitates Epiblast-Primitive Endoderm Fate Specification during Mouse Blastocyst Formation

doi: 10.1016/j.devcel.2019.10.011

Figure Lengend Snippet: PrE Specification and Spatial Segregation of ICM Lineages Is Impaired by Mechanical Inhibition of Lumen Expansion (A) Brightfield images of mechanical deflation. Magenta asterisk marks the needle tip. Dotted magenta line indicates lumen boundary. (B) Immunofluorescence images of EPI (Sox2) and PrE (Gata4) fate in pre-manipulation control (E3.5 WT), E4.0 post-manipulation control (E4.0 WT), and E4.0 mechanically inhibited (E4.0 Mechanical) embryos. Magenta arrowheads indicate the position of cells expressing high levels of Gata4 within the ICM. White dotted line indicates lumen boundaries. Average lumen volume in white text. Scale bars, 10 μm. (C) Boxplot of fluorescence levels of Sox2 (green) and Gata4 (magenta) in mechanically inhibited (Mech., N = 33) and post-manipulation control (WT, N = 28) E4.0 embryos. (D) Boxplot of spatial overlap between EPI and PrE lineages within post-manipulation control (WT, N = 27), mechanically inhibited (Mech., N = 33), E4.0 procedural control (Control, N = 11), and E4.0 simulation of complete overlap in WT conditions (Simulation, N = 27). ∗∗∗∗ p < 0.0001, ∗∗ p < 0.01. n.s., not significant. For boxplots: central mark indicates the median; lower edge, 25%; upper edge, 75%; lower whisker, Q1 − (1.5 × IQR), where IQR = Q3 − Q1; upper whisker, Q3 + (1.5 × IQR). See also Figure S6 and .

Article Snippet: The following primary antibodies were used in this study: rabbit anti-pERM (Cell Signaling, 3726), mouse anti-Cdx2 (BioGenex, MU392A-UC), goat anti-Sox2 (Santa Cruz Biotechnology, sc-17320), goat anti-Sox2 (R&D Systems, AF2018-SP), rabbit anti-Sox2 (Cell Signaling, 23064), rabbit anti Gata4 (Santa Cruz Biotechnology, sc-9053), goat anti-Gata4 (R&D Systems, AF2606-SP), rabbit anti-GFP (MBL, 598), mouse anti-Rab11 (BD Biosciences; 610656), rat anti-integrin-β1 (Merck Millipore, MAB1997), goat anti-biotinylated Gata4 (R&D Systems, BAF2606) mouse anti-Oct3/4 (Santa Cruz Biotechnology, sc-5279) and mouse anti-Hsp47 (Enzo Life Sciences, M16.10A1).

Techniques: Inhibition, Immunofluorescence, Control, Expressing, Fluorescence, Whisker Assay

Journal: Developmental Cell

Article Title: Lumen Expansion Facilitates Epiblast-Primitive Endoderm Fate Specification during Mouse Blastocyst Formation

doi: 10.1016/j.devcel.2019.10.011

Figure Lengend Snippet: Perturbation of FGF4 Signaling in the Lumen Impacts Molecular Specification of EPI and PrE Lineages (A) Immunofluorescence images of EPI (Sox2) and PrE (Gata4) fate in E3.5 post-FGF4 deposition (E3.5 FGF4 Inj.), E3.5 post-PD173074 deposition (E3.5 PD Inj.), and E3.5 post-PBS deposition (E3.5 PBS Inj.). White dotted line indicates lumen boundaries. Average lumen volume in white text. Scale bars, 10 μm. (B) Boxplot of fluorescence levels of Sox2 (green) and Gata4 (magenta) in E3.5 post-FGF4 deposition (FGF4 Inj., N = 24), E3.5 post-PD173074 deposition (PD Inj., N = 21), and E3.5 post-PBS deposition (PBS Inj., N = 13) embryos. (C) Boxplot of luminal volume in E3.5 post-FGF4 deposition (FGF4 Inj., N = 24), E3.5 post-PD173074 deposition (PD Inj., N = 21), and E3.5 post-PBS deposition (PBS Inj., N = 13). ∗∗∗∗ p < 0.0001, ∗∗ p < 0.01. n.s., not significant. For boxplots: central mark indicates the median; lower edge, 25%; upper edge, 75%; lower whisker, Q1 − (1.5 × IQR) where IQR = Q3 − Q1; upper whisker, Q3 + (1.5 × IQR). See also Figure S7 .

Article Snippet: The following primary antibodies were used in this study: rabbit anti-pERM (Cell Signaling, 3726), mouse anti-Cdx2 (BioGenex, MU392A-UC), goat anti-Sox2 (Santa Cruz Biotechnology, sc-17320), goat anti-Sox2 (R&D Systems, AF2018-SP), rabbit anti-Sox2 (Cell Signaling, 23064), rabbit anti Gata4 (Santa Cruz Biotechnology, sc-9053), goat anti-Gata4 (R&D Systems, AF2606-SP), rabbit anti-GFP (MBL, 598), mouse anti-Rab11 (BD Biosciences; 610656), rat anti-integrin-β1 (Merck Millipore, MAB1997), goat anti-biotinylated Gata4 (R&D Systems, BAF2606) mouse anti-Oct3/4 (Santa Cruz Biotechnology, sc-5279) and mouse anti-Hsp47 (Enzo Life Sciences, M16.10A1).

Techniques: Immunofluorescence, Fluorescence, Whisker Assay

Journal: Developmental Cell

Article Title: Lumen Expansion Facilitates Epiblast-Primitive Endoderm Fate Specification during Mouse Blastocyst Formation

doi: 10.1016/j.devcel.2019.10.011

Figure Lengend Snippet: Luminal Deposition of FGF4 Partially Rescues EPI-PrE Specification in ATP1-Inhibited Embryos (A) Immunofluorescence images of EPI (Sox2) and PrE (Gata4) fate in E3.5 post-FGF4 deposition and Atp1 inhibition (E3.5 FGF4 250 μM), E3.5 post-PBS deposition and Atp1 inhibition (E3.5 PBS 250 μM), and E3.5 control embryos (E3.5 DMSO). White dotted line indicates lumen boundaries. Average lumen volume in white text. Magenta arrowhead indicates cell with high Gata4 expression relative to neighboring cells. Scale bars, 10 μm. (B) Boxplot of fluorescence levels of Sox2 (green) and Gata4 (magenta) in E3.5 post-FGF4 deposition and Atp1 inhibition (250 μM FGF4 Inj., N = 42 embryos), E3.5 post-PBS deposition and Atp1 inhibition (250 μM PBS Inj., N = 12), and E3.5 control embryos (DMSO, N = 12). (C) Boxplot of luminal volume in E3.5 post-FGF4 deposition and Atp1 inhibition (250 μM FGF4 Inj., N = 49 embryos), E3.5 post-PBS deposition and Atp1 inhibition (250μM PBS Inj., N = 9), and E3.5 control embryos (DMSO, N = 38). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. n.s., not significant. For boxplots: central mark indicates the median; lower edge, 25%; upper edge, 75%; lower whisker, Q1 − (1.5 × IQR), where IQR = Q3 − Q1; upper whisker, Q3 + (1.5 × IQR).

Article Snippet: The following primary antibodies were used in this study: rabbit anti-pERM (Cell Signaling, 3726), mouse anti-Cdx2 (BioGenex, MU392A-UC), goat anti-Sox2 (Santa Cruz Biotechnology, sc-17320), goat anti-Sox2 (R&D Systems, AF2018-SP), rabbit anti-Sox2 (Cell Signaling, 23064), rabbit anti Gata4 (Santa Cruz Biotechnology, sc-9053), goat anti-Gata4 (R&D Systems, AF2606-SP), rabbit anti-GFP (MBL, 598), mouse anti-Rab11 (BD Biosciences; 610656), rat anti-integrin-β1 (Merck Millipore, MAB1997), goat anti-biotinylated Gata4 (R&D Systems, BAF2606) mouse anti-Oct3/4 (Santa Cruz Biotechnology, sc-5279) and mouse anti-Hsp47 (Enzo Life Sciences, M16.10A1).

Techniques: Immunofluorescence, Inhibition, Control, Expressing, Fluorescence, Whisker Assay

Journal: Developmental Cell

Article Title: Lumen Expansion Facilitates Epiblast-Primitive Endoderm Fate Specification during Mouse Blastocyst Formation

doi: 10.1016/j.devcel.2019.10.011

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used in this study: rabbit anti-pERM (Cell Signaling, 3726), mouse anti-Cdx2 (BioGenex, MU392A-UC), goat anti-Sox2 (Santa Cruz Biotechnology, sc-17320), goat anti-Sox2 (R&D Systems, AF2018-SP), rabbit anti-Sox2 (Cell Signaling, 23064), rabbit anti Gata4 (Santa Cruz Biotechnology, sc-9053), goat anti-Gata4 (R&D Systems, AF2606-SP), rabbit anti-GFP (MBL, 598), mouse anti-Rab11 (BD Biosciences; 610656), rat anti-integrin-β1 (Merck Millipore, MAB1997), goat anti-biotinylated Gata4 (R&D Systems, BAF2606) mouse anti-Oct3/4 (Santa Cruz Biotechnology, sc-5279) and mouse anti-Hsp47 (Enzo Life Sciences, M16.10A1).

Techniques: Recombinant, Software

Journal: Stem cell research

Article Title: Generation of two human iPSC lines with Exon 3 mutations in BCL2-Associated Athanogene 3 ( BAG3 ) from dilated cardiomyopathy patients

doi: 10.1016/j.scr.2023.103019

Figure Lengend Snippet: Reagents details.

Article Snippet: Differentiation Markers (Ectoderm) , Goat anti-OTX2 , 1:200 , R and D Systems Cat# AF1979, RRID: AB_2157172.

Techniques: Immunocytochemistry, Staining