Journal: bioRxiv

Article Title: KGF induces podosome formation via integrin-Erk1/2 signaling in human immortalized oral epithelial cells

doi: 10.1101/508416

Figure Lengend Snippet: KGF Mediated its Effects on HIOECs through Erk1/2 and Akt Signaling. (A and B) Western blot detection of the expression level of KGFR in HIOECs after KGF treatment, (C and D) the phosphorylation level of Jak2 and Stat3 after KGF stimulation for 15 min, and (E and F) the phosphorylation level of Erk1/2, p38, Jnk, and Akt after KGF stimulation for 15 min. n=3 independent experiments. Analysis of matrix protein expression in protein extracts from each group by ImageJ. (G) EdU labeling assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. (H) The quantification shown in the right graph indicates the mean ± SEM. (I) MTT assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. Data are expressed as the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: HIOECs at 3x10 4 cells/well were seeded in the 96-well plate and exposed to KGF (10 ng/mL; Proteintech) for 24 h. For MTT assays, the cells were incubated with 20 μL of MTT solution (5 mg/mL) at 37°C for 4 h. The supernatant was discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well.

Techniques: Western Blot, Expressing, Phospho-proteomics, Labeling, MTT Assay

Journal: The Journal of Clinical Investigation

Article Title: Targeting FAPα-expressing hepatic stellate cells overcomes resistance to antiangiogenics in colorectal cancer liver metastasis models

doi: 10.1172/JCI157399

Figure Lengend Snippet: ( A ) Differential proteomic analysis of tumor tissues obtained from bevacizumab-sensitive and -resistant HCT116 CRCLM xenografts (log 2 [fold change] > 1.0, P < 0.05; n = 3). ( B ) Immunohistochemical staining and quantification of FGFBP1 in the tumor-liver interface of CRCLM xenografts ( n = 6). Scale bar: 50 μm. ( C and D ) Western blotting ( C ) and ELISA ( D ) analysis of the expression of FGFBP1 in HCT116 and HT-29 cells isolated from the indicated CRCLM xenografts ( n = 3). ( E ) H&E staining of the tumor-liver interface of CRCLM xenografts. Scale bar: 100 μm. Quantification of RHGP is shown ( n = 6). ( F ) Immunofluorescence staining of the EpCAM + tumor cells (green) that infiltrated the liver parenchyma and hijacked the CD31 + sinusoidal blood vessels (red) in the tumor-liver interface of CRCLM xenografts. Scale bar: 20 μm. Quantification of the co-opted sinusoidal blood vessels is shown ( n = 6). ( G ) Immunofluorescence staining of FAPα + HSCs (green) attached to the CD31 + sinusoidal blood vessels (red) in the tumor-liver interface of CRCLM xenografts. Scale bar: 10 μm. Quantification of the co-opted FAPα + HSCs is shown ( n = 6). Dotted lines indicate the tumor-liver interface. LM, liver metastases; T, tumor; L, liver. Data are presented as mean ± SEM. NS, no significance. *** P < 0.001 (2-tailed, unpaired t test).

Article Snippet: An FGFBP1 ELISA Kit (Boster) and CXCL5 ELISA Kit (Solarbio) were used to measure FGFBP1 or CXCL5 levels in the cell culture supernatant according to the manufacturers’ instructions.

Techniques: Immunohistochemical staining, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Isolation, Immunofluorescence

Journal: The Journal of Clinical Investigation

Article Title: Targeting FAPα-expressing hepatic stellate cells overcomes resistance to antiangiogenics in colorectal cancer liver metastasis models

doi: 10.1172/JCI157399

Figure Lengend Snippet: ( A ) CT scan images of CRCLM patients with DHGP or PHGP treated preoperatively with Chemo or Bev+Chemo, and red arrows indicate the tumor lesions. Quantification of the morphologic response to RHGP is shown. ( B ) H&E staining of the tumor-liver interface of human CRCLM tissues. Quantification of RHGP is shown ( n = 6). Scale bar: 100 μm. ( C ) Immunohistochemical staining of FGFBP1 in the tumor-liver interface of human CRCLM tissues. Scale bar: 20 μm. Quantification of FGFBP1 staining is shown (right panel, n = 6). ( D ) Immunofluorescence staining of the EpCAM + cancer cells (green) that infiltrated the liver parenchyma and hijacked the CD31 + sinusoidal blood vessels (red), and FAPα (gray) expression in the co-opted HSCs in the tumor-liver interface of human CRCLM tissues. Scale bar: 20 μm. Quantification of the co-opted sinusoidal blood vessels and FAPα + HSCs is shown ( n = 6). Yellow arrows indicate the FAPα + HSCs. ( E ) Immunofluorescence staining of p-FGFR1(green) in HSCs attached to the CD31 + sinusoidal blood vessels (red) in the tumor-liver interface of human CRCLM tissues. Scale bar: 20 μm. White arrows indicate the p-FGFR1 + HSCs. ( F ) Pearson’s correlation analysis of FAPα + HSCs and the co-opted sinusoidal blood vessels, percentage of RHGP, or FGFBP1 expression in human CRCLM tissues ( n = 82). ( G ) CT scan images of CRCLM patients with RHGP treated preoperatively with Chemo or Bev+Chemo, and red arrows indicate the tumor lesions. Quantification of the change of tumor burden is shown. Dotted lines indicate the tumor-liver interface. Chemo, chemotherapy; Bev, bevacizumab; T, tumor; L, liver. Data are presented as mean ± SEM. NS, no significance. ** P < 0.01, *** P < 0.001 (2-tailed, unpaired t test).

Article Snippet: An FGFBP1 ELISA Kit (Boster) and CXCL5 ELISA Kit (Solarbio) were used to measure FGFBP1 or CXCL5 levels in the cell culture supernatant according to the manufacturers’ instructions.

Techniques: Computed Tomography, Staining, Immunohistochemical staining, Immunofluorescence, Expressing

Journal: Cells

Article Title: Regulation of Transplanted Cell Homing by FGF1 and PDGFB after Doxorubicin Myocardial Injury

doi: 10.3390/cells10112998

Figure Lengend Snippet: Doxorubicin increases FGF-1 and PDGF-B protein levels in adult heart ventricular cells. Mice were given intraperitoneal injections of saline (S) or doxorubicin (D) and their hearts were harvested after 24 h (Day 1) or 72 h (Day 3). Using Western blotting, lysates containing equal amounts of total protein from the ventricular tissue of the differently treated animals were tested for FGF-1 and PDGF-B protein levels. When standardized against GAPDH, the level of both proteins (FGF-1 and PDGF-B) was found to be significantly higher in the Dox-treated ventricles than in the saline treated ventricles by Day 3. * p < 0.05 vs. saline (S), One-way ANOVA with Tukey’s multiple comparisons test, N = 3 independent experiments for each group.

Article Snippet: For the analysis of growth factor effects, the wells contained 1% FBS (Low Serum) DMEM and 40 ng/mL of FGF-1 (rMuFGF1 TM , Gold Biotechnology, St. Louis, MO, USA), its neutralizing antibody (Santa Cruz sc-7910, 100 ng/mL), or both.

Techniques: Western Blot

Journal: Cells

Article Title: Regulation of Transplanted Cell Homing by FGF1 and PDGFB after Doxorubicin Myocardial Injury

doi: 10.3390/cells10112998

Figure Lengend Snippet: FGF-1 and PDGF-B increase embryonic ventricular cell migration, whereas their neutralizing antibodies decrease migration. 300 μL of 0.5 × 10 6 cells/mL serum-free cell suspensions were incubated for 22 h at 37 °C in polycarbonate membrane inserts with 8 μm pores, with surrounding wells containing 1% FBS DMEM and FGF-1, its antibody, or both ( A ) or PDGF-B, its antibody, or both ( B ). The inserts were then removed, migratory cells that had passed through the pores were washed and stained, and the cell stain was solubilized for measurement of absorbance. * p < 0.05 vs. all other groups; # p < 0.05 vs. all other groups, One-way ANOVA with Tukey’s multiple comparisons test, N = 3 independent experiments for each group.

Article Snippet: For the analysis of growth factor effects, the wells contained 1% FBS (Low Serum) DMEM and 40 ng/mL of FGF-1 (rMuFGF1 TM , Gold Biotechnology, St. Louis, MO, USA), its neutralizing antibody (Santa Cruz sc-7910, 100 ng/mL), or both.

Techniques: Migration, Incubation, Staining

Journal: Scientific Reports

Article Title: Myeloid Cell-Specific Lipin-1 Deficiency Stimulates Endocrine Adiponectin-FGF15 Axis and Ameliorates Ethanol-Induced Liver Injury in Mice

doi: 10.1038/srep34117

Figure Lengend Snippet: Wild-type (WT) or mLipin-1KO (KO) mice were pair-fed either a control diet or an ethanol-containing diet for 10 days followed by single gavage of ethanol. ( A ) Representative Western analysis of liver Ac-NF-κB, NF-κB, p-IκBα and IκBα. ( B ) Relative liver protein levels of liver Ac-NF-κB, NF-κB, p-IκBα and IκBα. ( C ) Male C57/6J mice were treated by intraperitoneal injection 1 mg/kg mouse body weight of recombinant FGF19 for six hours. Relative adipose mRNA levels of adiponectin. Relative liver mRNA of Cyp7a1. ( D ) Male intestinal FGF15 knock out (FGF15KO) mice and LOX littermates (WT) mice were fed chow diets. Relative adipose mRNA levels of adiponectin. Relative liver mRNA levels of Cyp7a1. ( E ) Male C57/6J mice were treated by intraperitoneal injection with 0.5 μg/g mouse body weight of recombinant human globular adiponectin (gAcrp) for three days. Relative ileum FGF15. Relative liver mRNA of Cyp7a1. Results are expressed as means ± SEM (n = 4–6 mice). Means without a common letter differ, P < 0.05.

Article Snippet: 12 week old male C57BL/6J mice were treated by intraperitoneal injection with 1 mg/kg mouse body weight of recombinant FGF19 (Gold Biotech, St. Louis, MO) in PBS or vehicle (PBS) and then sacrificed six hours later.

Techniques: Western Blot, Injection, Recombinant, Knock-Out