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: Cell reports

Article Title: Inhibition of FGF10-ERK signal activation suppresses intraductal papillary neoplasm of the bile duct and its associated carcinomas.

doi: 10.1016/j.celrep.2021.108772

Figure Lengend Snippet: Figure 5. Fgf10-induced IPNB shows stepwise carcinogenesis (A) Schematic representation of iFGF10: LSL-KrasG12D:Alb-Cre or Pdx1-Cre (iFGF10KA or iFGF10KP, respectively) mice. (B) H&E images of IPNB lesion in iFGF10KA (Kras+) and iFGF10A (Kras) mice following 0.002% Dox administration for 6 weeks (N = 5 each). Insets indicate high magnification of the rectangles.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Monoclonal Anti-Human Cytokeratin (clones AE1/AE3) DAKO Cat# M3515 LOT: 10066159 Rabbit Polyclonal Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Cell Signaling Technology Cat# 4370 RRID:AB_2315112 Rabbit polyclonal Anti-alpha smooth muscle Actin [1A4] Abcam Cat# ab7817 Rabbit polyclonal Anti-S100 antibody Abcam Cat# ab166649 Rabbit polyclonal Anti-FGF10 antibody (for mouse) Santa Cruz Cat# sc-293208 Rabbit polyclonal Anti-FGF10 antibody (for human) Abcam Cat# ab80064 Rabbit polyclonal Anti-FGF10 antibody Sigma-Aldrich Cat#ABN44 Mouse monoclonal Anti-MUC5AC antibody Abcam Cat# ab3649 Mouse monoclonal Anti-MUC6 antibody Leica Biosystems Cat# 130307 Rabbit polyclonal Anti-MUC2 antibody Novus Biologicals Cat# NBP1-31231 Rabbit monoclonal Anti-MUC1 antibody Invitrogen Cat# MA5-35250 Rabbit polyclonal Anti-Phospho-FGFR Pan (Tyr653, Tyr654) antibody Invitrogen Cat# PA5-105938 Rabbit polyclonal Anti-Src (phospho Y418) antibody Abcam Cat# ab4816 Rabbit Polyclonal RFP Antibody Preadsorbed Rockland Immunochemicals Cat# 600-401-379 Goat Anti-Mouse IgG H&L (Alexa Fluor 488) preadsorbed Abcam Cat# ab150117 Goat Anti-Rabbit IgG H&L (Alexa Fluor 594) preadsorbed Abcam Cat# ab150084 Rabbit IgG (Control Antibody) Vector Laboratories Cat# I-1000 Mouse IgG (Control Antibody) Vector Laboratories Cat# I-2000 Bacterial and virus strains Escherichia coli strain SW105 Frederick National Laboratory Cancer Research, NCI NA Biological samples Human biliary tumor samples This paper NA Chemicals, peptides, and recombinant proteins Tamoxifen Sigma Cat# T5648 Recommbinant mouse FGF10 R&D Systems Cat# 6224-FG Recommbinant human FGF10 R&D Systems Cat# 345-FG-025 thioacetamide Sigma Cat# 163678 Doxycycline hyclate Sigma Cat# D9891 Polyinosinic-Polycytidylic acid Sigma Cat# P1530 - Cellstain - DAPI solution DOJINDO Cat# D523 PD0325901 Selleck Chemicals Cat# S1036 L-arabinose Sigma Cat# A3256 Sucrose FUJIFILM Cat# 192-00017 dimethyl sulfoxide FUJIFILM Cat# 043-07211 (Continued on next page) Cell Reports 34, 108772, February 23, 2021 e1

Techniques:

Journal: Cell reports

Article Title: Inhibition of FGF10-ERK signal activation suppresses intraductal papillary neoplasm of the bile duct and its associated carcinomas.

doi: 10.1016/j.celrep.2021.108772

Figure Lengend Snippet: Figure 6. Fgf10 is required for maintaining papillary structure defined as IPNB (A) Experimental design of the withdrawal of Dox and/or TAA administration to iFGF10 mice: (1) Dox-only (Dox+) administration and (2) TAA+Dox+ administration followed by the withdrawal (OFF) (N = 5 each). ‘‘IPNB establishment time’’ and ‘‘endpoint time’’ are indicated.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Monoclonal Anti-Human Cytokeratin (clones AE1/AE3) DAKO Cat# M3515 LOT: 10066159 Rabbit Polyclonal Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Cell Signaling Technology Cat# 4370 RRID:AB_2315112 Rabbit polyclonal Anti-alpha smooth muscle Actin [1A4] Abcam Cat# ab7817 Rabbit polyclonal Anti-S100 antibody Abcam Cat# ab166649 Rabbit polyclonal Anti-FGF10 antibody (for mouse) Santa Cruz Cat# sc-293208 Rabbit polyclonal Anti-FGF10 antibody (for human) Abcam Cat# ab80064 Rabbit polyclonal Anti-FGF10 antibody Sigma-Aldrich Cat#ABN44 Mouse monoclonal Anti-MUC5AC antibody Abcam Cat# ab3649 Mouse monoclonal Anti-MUC6 antibody Leica Biosystems Cat# 130307 Rabbit polyclonal Anti-MUC2 antibody Novus Biologicals Cat# NBP1-31231 Rabbit monoclonal Anti-MUC1 antibody Invitrogen Cat# MA5-35250 Rabbit polyclonal Anti-Phospho-FGFR Pan (Tyr653, Tyr654) antibody Invitrogen Cat# PA5-105938 Rabbit polyclonal Anti-Src (phospho Y418) antibody Abcam Cat# ab4816 Rabbit Polyclonal RFP Antibody Preadsorbed Rockland Immunochemicals Cat# 600-401-379 Goat Anti-Mouse IgG H&L (Alexa Fluor 488) preadsorbed Abcam Cat# ab150117 Goat Anti-Rabbit IgG H&L (Alexa Fluor 594) preadsorbed Abcam Cat# ab150084 Rabbit IgG (Control Antibody) Vector Laboratories Cat# I-1000 Mouse IgG (Control Antibody) Vector Laboratories Cat# I-2000 Bacterial and virus strains Escherichia coli strain SW105 Frederick National Laboratory Cancer Research, NCI NA Biological samples Human biliary tumor samples This paper NA Chemicals, peptides, and recombinant proteins Tamoxifen Sigma Cat# T5648 Recommbinant mouse FGF10 R&D Systems Cat# 6224-FG Recommbinant human FGF10 R&D Systems Cat# 345-FG-025 thioacetamide Sigma Cat# 163678 Doxycycline hyclate Sigma Cat# D9891 Polyinosinic-Polycytidylic acid Sigma Cat# P1530 - Cellstain - DAPI solution DOJINDO Cat# D523 PD0325901 Selleck Chemicals Cat# S1036 L-arabinose Sigma Cat# A3256 Sucrose FUJIFILM Cat# 192-00017 dimethyl sulfoxide FUJIFILM Cat# 043-07211 (Continued on next page) Cell Reports 34, 108772, February 23, 2021 e1

Techniques:

Journal: British Journal of Cancer

Article Title: FGF10/FGFR2 signal induces cell migration and invasion in pancreatic cancer

doi: 10.1038/sj.bjc.6604473

Figure Lengend Snippet: Expression patterns of FGFR2 and FGF10 in normal pancreas and pancreatic cancer. The magnification is shown in the right bottom corner of each figure. ( A and B ) Immunostaining of FGFR2 ( A ) and FGF10 ( B ) in normal pancreas, showing that FGFR2 is expressed weakly in ductal cells ( A , arrow) and acinar cells ( A , arrow head), and that no obvious FGF10 staining is found in normal pancreatic tissue, including ductal cells ( B , arrows). ( C and D ) Immunostaining of FGFR2 ( C ) and FGF10 ( D ) in pancreatic cancer tissues, showing that FGFR2 is expressed in cancer cells ( C ), whereas FGF10 is expressed in scattered cells in the stroma surrounding cancer cells ( D , arrows). ( E and F ) Immunostaining of FGFR2 in pancreatic cancer cells. ( E ) Representative result from the FGFR2 high expression group, indicating higher FGFR2 expression in cancer cells (arrows) compared with islets (arrow head). ( F ) Representative result from the FGFR2 low expression group, showing lower FGFR2 expression in cancer cells (arrows) compared with islet (arrow heads). ( G and H ) Immunostaining of FGF10 ( G ) and CD3 ( H ), marker for T cell. Fibroblast growth factor 10 and CD3 are both expressed in scattered cells with similar cell shape in the stroma surrounding cancer cells (arrows).

Article Snippet: They were then rinsed and blocked in 10% H 2 O 2 solution with methanol for 10 min. Next, the sections were incubated with goat anti-human FGF10 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) at 1 : 20 dilution, mouse anti-human FGFR2 monoclonal antibody (R&D Systems) at 1 : 10 dilution, or rabbit anti-human CD3 monoclonal antibody (ready-to-use without dilution) (Thermo Fisher Scientific Anatomical Pathology, Fremont, CA, USA) overnight at 4°C.

Techniques: Expressing, Immunostaining, Staining, Marker

Journal: British Journal of Cancer

Article Title: FGF10/FGFR2 signal induces cell migration and invasion in pancreatic cancer

doi: 10.1038/sj.bjc.6604473

Figure Lengend Snippet: Fibroblast growth factor 10 induces cell migration and invasion in pancreatic cell lines with FGFR2-IIIb expression. ( A ) RT–PCR analysis of FGF10 and FGFR2-IIIb in four pancreatic cell lines and cDNA obtained from normal lung tissue as a positive control. None of the cell lines express FGF10. MIA PaCa-2 and PanC-1 cells do not express FGFR2-IIIb, but CFPAC-1 and AsPC-1 do express this gene. ( B ) Representative results of cell migration (upper panels) and invasion (lower panels) for CFPAC-1 cells. Representative migrated and invaded cells are indicated with arrows. ( C and D ) Cell migration ( C ) and invasion ( D ) assay of all four cell lines cultured without (white column) or with (black column) FGF10 (100 ng ml −1 ). FGF10-induced cell migration and invasion in CFPAC-1 and AsPC-1 cells, but not in MIAPaCa-2 and PanC-1 cells. The numbers of migrated or invaded cells cultured with FGF10 are shown relative to a value of 100% for cell migration without ligand. ( E and F ) Inhibition of FGFR2-IIIb signalling by an FGFR2-IIIb/IgG chimera in CFPAC-1 cells. Migration ( E ) and invasion ( F ) assay. The numbers of migrated or invaded cells are shown relative to a value of 100% for cells cultured without FGF10 or chimera (control; white column). FGF10-induced migration and invasion in CFPAC-1 cells (black column). Addition of the FGFR2-IIIb/IgG chimera completely eliminated the effects of FGF10 (grey column), whereas the chimera itself did not affect cell migration and invasion of CFPAC-1 cells (striped column). * P <0.05.

Article Snippet: They were then rinsed and blocked in 10% H 2 O 2 solution with methanol for 10 min. Next, the sections were incubated with goat anti-human FGF10 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) at 1 : 20 dilution, mouse anti-human FGFR2 monoclonal antibody (R&D Systems) at 1 : 10 dilution, or rabbit anti-human CD3 monoclonal antibody (ready-to-use without dilution) (Thermo Fisher Scientific Anatomical Pathology, Fremont, CA, USA) overnight at 4°C.

Techniques: Migration, Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Cell Culture, Inhibition, Control

Journal: British Journal of Cancer

Article Title: FGF10/FGFR2 signal induces cell migration and invasion in pancreatic cancer

doi: 10.1038/sj.bjc.6604473

Figure Lengend Snippet: Fibroblast growth factor 10 induces expression of mRNA for MT1-MMP and TGF- β 1 in CFPAC-1 cells. The figure shows the relative copy numbers of MT1-MMP ( A ) and TGF- β 1 ( B ) mRNA in CFPAC-1 cells cultured with 100 ng ml −1 of FGF10 for the indicated times. The MT1-MMP and TGF β 1 mRNA/GAPDH mRNA copy number ratios are shown relative to those of cells without FGF10 stimulation (0 h). The concentration of TGF- β 1 protein in medium also increased in time-dependent manner in CFPAC-1 cells 48 h after addition of FGF10 (100 ng ml −1 ) ( C ). FGF receptor-2-IIIb/IgG chimera (500 ng ml −1 ) inhibited this TGF- β 1 secretion by FGF10, whereas chimera alone did not affect TGF- β 1 secretion by itself ( D ). * P <0.05; NS=not significant.

Article Snippet: They were then rinsed and blocked in 10% H 2 O 2 solution with methanol for 10 min. Next, the sections were incubated with goat anti-human FGF10 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) at 1 : 20 dilution, mouse anti-human FGFR2 monoclonal antibody (R&D Systems) at 1 : 10 dilution, or rabbit anti-human CD3 monoclonal antibody (ready-to-use without dilution) (Thermo Fisher Scientific Anatomical Pathology, Fremont, CA, USA) overnight at 4°C.

Techniques: Expressing, Cell Culture, Concentration Assay