Journal: Developmental biology

Article Title: Retinoic acid selectively regulates Fgf10 expression and maintains cell identity in the prospective lung field of the developing foregut.

doi: 10.1016/j.ydbio.2004.04.039

Figure Lengend Snippet: Fig. 4. Whole mount ISH of Fgfr2b and Fgf10 mRNA in vivo and in foregut cultures. (A) Fgfr2b mRNA is expressed throughout the foregut endoderm including the respiratory tract in E9.5 embryos and in (B) control cultures. (B, C) Fgfr2b expression is not disrupted by BMS treatment. (D) At E8.5, Fgf10 is expressed in the mesoderm of thyroid; subsequently (E9.5), signals appear in lung and thyroid domains. (E) In control cultures, these domains are preserved. (F) BMS treatment of explants younger than the 15-somite stage, but not older than that (G), prevents Fgf10 mRNA expression in the presumptive respiratory mesoderm (*). RA (106 M) has no obvious effects on Fgf10 expression. Area between dotted lines (in yellow) corresponds to the region of disrupted Fgf10 expression. Scale bars in A and E represent 85 and 150 Am, respectively.

Article Snippet: In some experiments, heparin beads soaked in human recombinant FGF10 (100 ng/Al, R&D Systems) or PBS (buffer) were grafted onto foreguts after 48 h culture.

Techniques: In Vivo, Control, Expressing

Journal: Developmental biology

Article Title: Retinoic acid selectively regulates Fgf10 expression and maintains cell identity in the prospective lung field of the developing foregut.

doi: 10.1016/j.ydbio.2004.04.039

Figure Lengend Snippet: Fig. 6. Exogenous FGF10 rescues lung budding and epithelial differentiation in BMS-treated cultures. PCNA staining in sagittal sections (A, D) and whole mount ISH of Ttf1 (B, E) and Sp-C (C, F) in 6-day cultures engrafted with heparin beads (human recombinant FGF10, PBS buffer). (A) FGF10 beads induced lung bud formation, local PCNA labeling and expression of Ttf1 (B) and Sp-C mRNA (C) in BMS-treated explants. PBS beads failed to induce a local increase in proliferation (D) and had no detectable Ttf1 (E) or Sp-C (F) signals in the respiratory region of foregut. Note the preserved Ttf1 staining in thyroid (B, E). (G–J) Quantitative analysis of PCNA- and TUNEL-stained sections from BMS-treated explants in the lung field. (H) Engraftment of an FGF10 bead results in a significant increase in the relative number of PCNA-labeled cells in the adjacent endoderm (G) as compared to a PBS bead. Mesodermal PCNA (G) or TUNEL labeling (I) in endoderm or mesoderm is not altered by FGF10. Graphs (H, J) represent mean and standard error; *P < 0.05. Lu, lung; Th, thyroid. Scale bar in E represents 240 Am.

Article Snippet: In some experiments, heparin beads soaked in human recombinant FGF10 (100 ng/Al, R&D Systems) or PBS (buffer) were grafted onto foreguts after 48 h culture.

Techniques: Staining, Recombinant, Labeling, Expressing, TUNEL Assay

Journal: Developmental biology

Article Title: Retinoic acid selectively regulates Fgf10 expression and maintains cell identity in the prospective lung field of the developing foregut.

doi: 10.1016/j.ydbio.2004.04.039

Figure Lengend Snippet: Fig. 7. Lung agenesis and disrupted expression of Ttf1 protein and Fgf10 mRNA in the respiratory region of foregut of vitamin A-deficient rats (A–D). H&E and Ttf1 immunostaining of transverse sections of control RAS foregut demonstrates (A) ongoing tracheoesophageal separation with Ttf1 expression restricted to the ventrally located tracheal epithelium and, more caudally, (B) primary lung bud epithelium intensely labeled with Ttf1. (C) In RAD foregut, the tube is irregularly shaped (*) and shows low levels of Ttf1 expression in the ventral endoderm at the prospective respiratory region that failed to bud. (D) Strong Ttf1 signals, however, are present in the thyroid bud. (E, F) Isotopic ISH of Fgf10 mRNA in transverse sections at the level of the lung buds in control RAS demonstrates localized signal in the mesoderm of the lung (F, enlarged), limb and urogenital tract (ug). In RAD (G, H), Fgf10 expression is preserved in the limb and urogenital tract but is disrupted in the presumptive lung field of the foregut (H, high magnification). Es, esophagus; fl, forelimb; hl, hindlimb; Lu, lung; Th, thyroid; Tr, trachea; RAD, RA deficient; RAS, RA sufficient; ug, urogenital ridge. Dashed boxes in E and G delineate close-up views in F and H, respectively. Arrowheads represent signals; asterisks indicate areas that failed to bud or to express Fgf10. Scale bars in B and G represent 80 and 480 Am, respectively.

Article Snippet: In some experiments, heparin beads soaked in human recombinant FGF10 (100 ng/Al, R&D Systems) or PBS (buffer) were grafted onto foreguts after 48 h culture.

Techniques: Expressing, Immunostaining, Control, Labeling

Journal: Developmental biology

Article Title: Retinoic acid selectively regulates Fgf10 expression and maintains cell identity in the prospective lung field of the developing foregut.

doi: 10.1016/j.ydbio.2004.04.039

Figure Lengend Snippet: Fig. 8. Proposed model for the role of RA in early lung morphogenesis. Early in mouse foregut organogenesis, RA signaling is active in all layers. (A) By the 15-somite stage, RA has induced proliferation of a critical mass of mesodermal cells in the lung field, while signaling in endoderm maintains identity in lung progenitor cells (in red). (B) At around 25 somites, the condensing mesodermal cells begin to locally express Fgf10 (in green), which then activates the Fgf pathway in the endoderm to expand the population of lung progenitor cells into a primary endodermal bud. (C) Once secondary buds form and branching morphogenesis starts, RA signaling is downregulated in the lung and Fgf10 expression becomes independent of RA.

Article Snippet: In some experiments, heparin beads soaked in human recombinant FGF10 (100 ng/Al, R&D Systems) or PBS (buffer) were grafted onto foreguts after 48 h culture.

Techniques: Expressing

Journal: Developmental biology

Article Title: Dermal fibroblast-derived growth factors restore the ability of beta(1) integrin-deficient embryonal stem cells to differentiate into keratinocytes.

doi: 10.1006/dbio.2000.0149

Figure Lengend Snippet: FIG. 5. Effects of specific growth factors on ES cell differentiation and detection of growth factors produced by HDF. Differentiation of wild-type (a) and b1-null (b) ES cells in the presence or absence (co, control) of various growth factors: human acidic and basic FGF (at final concentration of 50 and 10 ng/ml, respectively), KGF (100 ng/ml), FGF10 (20 ng/ml), and TGFa (25 ng/ml). Data from a single experiment are shown. (c) Quantitation of KGF levels in medium conditioned by HDF for 2–4 days. Data are means 6 SD of three separate experiments. (d, e) immunohistochemical staining of HDF with antibody to FGF10 (d) or second antibody alone (e). Scale bar: 50 mm.

Article Snippet: FGF10 was detected in HDF cultures by immunohistochemistry f paraformaldehyde-fixed cells, using goat anti-human FGF10 ntibody (R&D Systems) and an avidin-biotin horseradish peroxiase detection system (Vectastain ABC kit; Vector Laboratories, urlingame, CA).

Techniques: Cell Differentiation, Produced, Control, Concentration Assay, Quantitation Assay, Immunohistochemical staining, Staining

Journal: Molecular and Cellular Biology

Article Title: The RNA-Binding Protein Elavl1/HuR Is Essential for Placental Branching Morphogenesis and Embryonic Development

doi: 10.1128/mcb.01393-08

Figure Lengend Snippet: FIG. 10. Differential effects of HuR’s loss on the transcription and translation of its target mRNAs. (A) Densitometric quantitation of nuclei run-on experiments with radiolabeled RNAs from control and mutant MEFs hybridized to cold probes spotted onto nylon filters. Data (phosphorimager units standard deviations) were derived from three individual experiments. The asterisk denotes a significant difference with a P value of 0.01. (B) qRT-PCR detection of HuR target RNAs in monosomal (Mon)/polysomal (Pol) fractions from control and mutant MEFs. Data are derived from measurements in pooled fractions normalized to 2 mRNA and presented as percentages ( standard errors of the means) of total cytoplasmic mRNA. An asterisk denotes a significant difference with a P value of 0.05. (C) Detection and quantitation of immunoblotted proteins in total (for Ets2, FGF10, and Hoxd13) or nuclear (for Tbx4) extracts derived from control and mutant MEFs. Actin is shown as a loading control. Protein levels (phosphorimager units standard deviations from three independent samples) were normalized to actin. An asterisk denotes a significant difference with a P value of 0.05. (D) Detection of Ets2 (left) or Hoxd13 (right) in extracts from E10.5 placentas or E12.5 limbs, respectively. For Ets2, E10.5 embryonic extracts are shown as negative controls. Actin is shown as a loading control.

Article Snippet: The primary antibodies used were 3A2 and T17 for HuR, C-11 for actin, H-40 for Hoxd13, C-20 for Ets-2, H-121 for FGF10, and N-19 for TBX4 and were purchased from SantaCruz Biotech.

Techniques: Quantitation Assay, Control, Mutagenesis, Derivative Assay, Quantitative RT-PCR