primary antibodies against fgfr2  (Proteintech)

Journal: Pharmaceuticals

Article Title: Inhibition of FGFR2 Signaling by Cynaroside Attenuates Liver Fibrosis

doi: 10.3390/ph16040548

Figure Lengend Snippet: High expression of fibroblast growth factor receptor 2 (FGFR2) coincides with liver fibrosis. ( A ) The expression of FGFR2 in fibrotic liver and normal liver tissues was evaluated through data mining of human samples affected by Hepatitis B infection (GSE38941), alcohol abuse (GSE28619), and nonalcoholic steatohepatitis (GSE48452), as well as mouse samples induced with carbon tetrachloride (CCl 4 ) (GSE152329, GSE98577). The differential fold expression was determined by comparing the expression of FGFR2 in the fibrotic liver and normal liver tissues. ( B ) A comparison of changes in FGFR2 expression in liver fibrosis patients in remission (improved) or not in remission (not improved) with mining data of GSE175448. ( C ) The correlation between FGFR2 expression and the liver fibrosis score (Ishak score) was analyzed by comparing FGFR2 expression in three pairs of liver fibrotic tissues and normal liver tissues through immunohistochemistry. ( D ) FGFR2 expression in liver fibrosis tissues and normal liver tissues was verified by Western blot and ( E ) q-PCR analyses. ( E ) Liver tissues from mice treated with CCl 4 for different durations were used to determine the expression of the liver fibrosis markers Actin Alpha 2 (ACTA2) and FGFR2 by q-PCR and the trends of their expression with increasing days of induction. ( F ) The expression of ACTA2 and ( G ) FGFR2 was determined by q-PCR in liver tissues from mice treated with CCl 4 for varying durations. The statistical significance was determined based on the p -value. The results are marked as significant “*” when p < 0.05, “**” when p < 0.01, “***” when p < 0.001, and not significant (ns) if p ≥ 0.05.

Article Snippet: The Western blot and IHC analyses were performed using primary antibodies against FGFR2 (23328S, CST, Danvers, MA, USA), phospho-FGFRs (AF3285, R&D, Minneapolis, MN, USA), α-SMA (ab7817, Abcam, Waltham, MA, USA), FAP (AF3715, R&D, Minneapolis, MN, USA), and β-actin (MAB8929, R&D, Minneapolis, MN, USA).

Techniques: Expressing, Infection, Comparison, Immunohistochemistry, Western Blot, IF-P

Journal: Pharmaceuticals

Article Title: Inhibition of FGFR2 Signaling by Cynaroside Attenuates Liver Fibrosis

doi: 10.3390/ph16040548

Figure Lengend Snippet: FGFR2 drives the process of liver fibrosis. ( A ) The expression of liver fibrosis markers (ACTA2, fibroblast activation protein alpha (FAP), alpha-1 type I collagen (COL1A1)) was measured by qPCR. The samples were divided into four groups according to whether cells were induced with or without TGF-β and whether cells were overexpressed by FGFR2. ( B ) The expression of markers in wild-type and FGFR2-OE cell lines under TGF-β induction was analyzed by Western blot analysis. ( C ) The expression levels of α-SMA and ( D ) collagen secretion in LX-2 and Huh-7 were evaluated, when FGFR2 was overexpressed or knocked down upon equal TGF-β induction. ( E ) Cells were collected after 48 h of co-culture, and the expression of α-SMA and ( F ) type I collagen in the lower layer cells was determined by ELISA. The results are marked as significant “*” when p < 0.05, “**” when p < 0.01, and not significant (ns) if p ≥ 0.05.

Article Snippet: The Western blot and IHC analyses were performed using primary antibodies against FGFR2 (23328S, CST, Danvers, MA, USA), phospho-FGFRs (AF3285, R&D, Minneapolis, MN, USA), α-SMA (ab7817, Abcam, Waltham, MA, USA), FAP (AF3715, R&D, Minneapolis, MN, USA), and β-actin (MAB8929, R&D, Minneapolis, MN, USA).

Techniques: Expressing, Activation Assay, Western Blot, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, IF-P

Journal: Pharmaceuticals

Article Title: Inhibition of FGFR2 Signaling by Cynaroside Attenuates Liver Fibrosis

doi: 10.3390/ph16040548

Figure Lengend Snippet: CYN blocks the activity of FGFR2. ( A ) A total of 2081 natural compounds were screened through high-throughput affinity analysis chips, resulting in the identification of compounds capable of binding to the FGFR2 kinase domain. The affinity and p -value of each compound were represented, and the candidate compound CYN was located in the high-affinity region. ( B ) The binding signal of FGFR2 and CYN was analyzed using surface plasmon resonance (SPR) technology at five different concentrations, and the affinity constant K D was calculated by fitting the curve algorithm. ( C ) An in vitro FGFR2 catalytic system was established, and the kinase activity was compared by measuring the phosphorylation product generation and ( D ) ATP consumption during the catalytic process. A series of concentration gradients were set up to evaluate the correlation between the inhibitory effect of CYN and its concentration. ( E ) Computational docking was utilized to predict the binding site of CYN on FGFR2. The surface charges on the model are depicted in blue (negative) to red (positive), and the appearance of the compound CYN is represented in green. Noncovalent amino acids that provide binding for the compound are marked in red on the right figure. ( F ) The diagram depicts the binding of the CYN and FGFR2 amino acid residues. ( G ) Molecular dynamics (MD)simulations were conducted to compare the stability of FGFR2 protein and FGFR2-CYN complex in aqueous environments. The mean value of the root-mean-square deviation (RMSD)at the plateau period is marked by a dotted line. ( H ) SPR was used to measure the affinity between the CYN and FGFR2 proteins with single-point mutations. The affinity is ranked from strong to weak in the legend, and the right-side values represent the corresponding K D values. A higher K D value represents weaker binding of the mutated FGFR2 protein with CYN, indicating that CYN’s binding is more dependent on the mutated amino acid. The results are marked as significant “*” when p < 0.05, “**” when p < 0.01, “***” when p < 0.001, and not significant (ns) if p ≥ 0.05.

Article Snippet: The Western blot and IHC analyses were performed using primary antibodies against FGFR2 (23328S, CST, Danvers, MA, USA), phospho-FGFRs (AF3285, R&D, Minneapolis, MN, USA), α-SMA (ab7817, Abcam, Waltham, MA, USA), FAP (AF3715, R&D, Minneapolis, MN, USA), and β-actin (MAB8929, R&D, Minneapolis, MN, USA).

Techniques: Activity Assay, High Throughput Screening Assay, Binding Assay, SPR Assay, In Vitro, Phospho-proteomics, Concentration Assay, IF-P

Journal: Pharmaceuticals

Article Title: Inhibition of FGFR2 Signaling by Cynaroside Attenuates Liver Fibrosis

doi: 10.3390/ph16040548

Figure Lengend Snippet: CYN blocks the activation and development of liver fibrosis in vitro. ( A ) Evaluation of the inhibitory effect of CYN on the fibrosis-promoting effect of FGFR2. The fibrotic transformation of wild-type and FGFR2-OE LX-2 cells and Huh-7 cells was induced through TGF-β activation, followed by an intervention with CYN for the relevant groups. The expression changes of the fibrosis markers ACTA2 and COL1A1 were analyzed by qPCR. ( B ) Wild-type cell lines were employed in the aforementioned experiments, and the activation of FGFR2 was triggered by supplementation with the exogenous basic fibroblast growth factor (bFGF) factor. ( C ) Analysis of the extent of antagonism of CYN towards TGF-β Signaling. Activation induction models were established by adding or not adding TGF-β to the cell culture environment with or without the CYN intervention, and the expression of liver fibrosis markers was determined by qPCR, ( D ) Western blot, and ( E , F ) ELISA analyses. ( G ) A co-culture model was used to evaluate the blocking effect of CYN on liver fibrosis activation of signaling transmission. The activation intensity of lower-layer wild-type cells was measured and compared using α-SMA expression and collagen secretion. The results are marked as significant “*” when p < 0.05, “**” when p < 0.01, and not significant (ns) if p ≥ 0.05.

Article Snippet: The Western blot and IHC analyses were performed using primary antibodies against FGFR2 (23328S, CST, Danvers, MA, USA), phospho-FGFRs (AF3285, R&D, Minneapolis, MN, USA), α-SMA (ab7817, Abcam, Waltham, MA, USA), FAP (AF3715, R&D, Minneapolis, MN, USA), and β-actin (MAB8929, R&D, Minneapolis, MN, USA).

Techniques: Activation Assay, In Vitro, Transformation Assay, Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Blocking Assay, Transmission Assay, IF-P

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 1. Expression of FGFR2 in CRC. (A) Higher FGFR2 expression and lower FGFR2 expression in human CRC tissues and adjacent normal tissues (original magnification 3200). (B) FGFR2 IHC staining of cancer tissues and corresponding adjacent normal tissues (original magnification 3200). (C) H-score of FGFR2 expression of cancer tissues and adjacent normal tissues (D) Kaplan–Meier survival curves of CRC patients with FGFR2 high expression and FGFR2 low expression. Tumor tissues with H-scores greater than the median of all scored tumor tissues were classified as high FGFR2 expression. Data were presented as the mean 6 SEM. ***p , 0.001.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Expressing, Immunohistochemistry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 2. Expression of PD-L1 in CRC. (A) Higher PD-L1 expression and lower PD-L1 expression in human CRC tissues and adjacent normal tissues (original magnification 3200). (B) Representative PD-L1 IHC staining of cancer tissues and corresponding adjacent normal tissues (original magnification 3200). (C) H-score of PD-L1 expression of cancer tissues and adjacent normal tissues. (D) Kaplan–Meier survival curves of CRC patients with PD-L1 high expression and PD-L1 low expression. Tumor tissues with H-scores greater than the median of all scored tumor tissues were classified as high PD-L1 expression. (E) The costaining of PD-L1 and FGFR2 in CRC tissue (original magnification 3400). Data were presented as the mean 6 SEM. ***p , 0.001.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Expressing, Immunohistochemistry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 3. Correlation of FGFR2 and PD-L1. (A) Correlation analysis of FGFR2 and PD-L1 expression of tissue microarray. (B) Correlation analysis of FGFR2 and PD-L1 gene expression in the TCGA cohort. (C) Correlation analysis of FGFR2 and PD-L1 gene expression of N2 CRC patients in the TCGA cohort. (D) Correlation analysis of FGFR2 and PD-L1 gene expression of clinical stage III CRC patients in the TCGA cohort.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Expressing, Microarray, Gene Expression

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 4. PD-L1 expression is correlated with the FGF7/FGFR2 signaling pathway in vitro. (A) PD-L1 expression and FGFR2 expression in four CRC cell lines and one gastric cancer cell line were analyzed by Western blotting. GAPDH was an internal control. (B) The expression of FGFR2 and PD-L1 in NCI-H716 cells transduced with lentivirus was examined by Western blot. Ctrl: normal NCI-H716 cells; Vector: NCI-H716 cells transduced with empty vector lentivirus; sh1-3: NCI-H716 cells transduced with FGFR2-knockdown lentivirus. (C) The expression of FGFR2 and PD-L1 in SW480 cells transduced with lentivirus was examined by Western blot. Ctrl: normal SW480 cells; Vector: SW480 cells transduced with empty vector lentivirus; FGFR2: SW480 cells transduced with FGFR2-overexpression lentivirus. (D) The relative expression of FGFR2 and PD-L1 mRNA in transduced cells and normal SW480 cells are detected by quantitative RT-PCR. (E and F) Cells were incubated with different concentrations (0, 10, and 50 ng/ml) of FGF7 for 24 h, and the expression of FGFR2, phospho-FGFR, and PD-L1 was evaluated by Western blot. Cells were incubated with FGF7 (10 ng/ml) for the indicated times and the expression of FGFR2, phospho-FGFR, and PD-L1 was evaluated by Western blot. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Expressing, In Vitro, Western Blot, Control, Transduction, Plasmid Preparation, Knockdown, Over Expression, Quantitative RT-PCR, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 5. FGF7/FGFR2 upregulates PD-L1 through the JAK/STAT3 signaling pathway. (A) Cells were pretreated with FGF7 (10 ng/ml, 48 h), and PD-L1 was assessed by Western blot after being treated with different concentrations of LY294002, U0126, SB203580, SP600125, RAD001, and AG490 for 24 h. (B) The expression of JAK2, phospho-JAK2, STAT3, and phospho-STAT3 was detected by Western blot after SW480 and NCI-H716 cells were incubated with FGF7 (10 ng/ml) for the indicated times. (C) The expression of JAK2, phospho-JAK2, STAT3, and phospho-STAT3 were detected in normal cells, FGFR2-overexpression cells, and FGFR2 knockdown cells. **p , 0.01.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Western Blot, Expressing, Incubation, Over Expression, Knockdown

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 6. FGFR2-induced upregulation of PD-L1 in human CRC cells promoted the apoptosis of Jurkat T cells. Jurkat cells were cocultured with SW480-vector cells, SW480-FGFR2 cells, and SW480-FGFR2 cells pretreated with AG490 (tumor cell to Jurkat cell ratio equals 10:1) for 24 h. Extent of apoptosis in Jurkat cells was determined by flow cytometry using FITC–annexin V. The experiment was repeated three times. **p , 0.01.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Plasmid Preparation, Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: FGFR2 Promotes Expression of PD-L1 in Colorectal Cancer via the JAK/STAT3 Signaling Pathway.

doi: 10.4049/jimmunol.1801199

Figure Lengend Snippet: FIGURE 7. AG490 inhibits tumor growth and the expression of PD-L1 in xenograft model. Mice were randomly divided into three groups (SW480 cells with vector group, FGFR2 group, FGFR2 plus AG490 group). AG490 (0.5 mg/mouse/d) and DMSO (20 ml/mouse/d) were administered by i.p. injection. (A) s.c. tumor growth was monitored for 21 d. The growth of tumor volume of each group was shown as a line chart. (B) IHC analyses of FGFR2 and PD-L1 from tumor samples in each group were shown (original magnification 3200). (C) FGF7 induces the activation of FGFR2 accompanied by the upregulation of PD-L1 expression as a result of JAK/STAT3 signaling pathway activation. PD-L1 binds to T cell–derived PD-1 to promote tumor immunosuppression and CRC progression. Data were presented as mean 6 SEM. *p , 0.05.

Article Snippet: Specific primary Abs against FGFR2 (1:100, TA503137; OriGene, Rockville, MD) and PD-L1 (1:100, ab205921; Abcam, Cambridge, U.K.) were used for IHC.

Techniques: Expressing, Plasmid Preparation, Injection, Activation Assay, Derivative Assay

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Generation of the mice with fibroblast-specific deletion of FGFR2. a Genotyping of the mice by PCR assay of genomic DNA. Lane 1: genotype FGFR2 fl/fl, Cre−/−; lane 2: genotype FGFR2 fl/wt, Cre−/−; lane 3: genotype FGFR2 fl/fl, Cre+/−, designated as S100A4-FGFR2−/−. b Representative immunohistochemical staining images showing the loss of FGFR2 in interstitial cells from the fibrotic kidneys in S100A4-FGFR2−/− mice. c-i Mice with fibroblast-specific ablation of FGFR2 exhibited little difference in body weight (c), serum creatinine (d), serum BUN (e), kidney weight index (f), urinary NAG (g), urinary lysozyme (h), and urinary albumin (i) levels compared to their control littermates within 8 weeks after birth (n = 3). j Representative images for PAS and H&E staining showing no obvious kidney morphological abnormality in S100A4-FGFR2−/− mice at 8 weeks after birth. BUN, blood urea nitrogen; FGFR, fibroblast growth factor receptor; NAG, N-acetyl-β-d-glucosaminidase; Scr, serum creatinine; Ucr, urinary creatinine.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Deletion of FGFR2 in fibroblasts ameliorated kidney fibrosis after IRI. a Representative images for PAS and Sirius Red staining in sham and IRI kidneys from S100A4-FGFR2−/− mice and control littermates. b Graph showing quantitative results for the area staining positive for Sirius Red. * p < 0.05 versus control littermates, n = 6. c Graph showing the results for total collagen content in kidneys from S100A4-FGFR2−/− mice and control littermates. * p < 0.05 versus control littermates, n = 6. FGFR, fibroblast growth factor receptor; IRI, ischemia/reperfusion injury.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: Staining, Control

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Deletion of FGFR2 in fibroblasts inhibits FN and α-SMA expression in the kidneys after IRI. a Representative images for FN and α-SMA immunostaining in the kidneys from S100A4- FGFR2−/− mice and control littermates. b Western blot assay showing the abundance of FN and α-SMA in the kidneys from S100A4-FGFR2−/− mice and control littermates. Numbers indicate individual animals. The gels were run under the same experimental conditions. c, d Graphs showing the semiquantitative results for the abundance of α-SMA (c) and FN (d). * p < 0.05 versus control littermates, n = 4. α-SMA, alpha smooth muscle actin; FGFR, fibroblast growth factor receptor; FN, fibronectin; IRI, ischemia/reperfusion injury.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: Expressing, Immunostaining, Control, Western Blot

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Interstitial cell proliferation was diminished in the IRI kidneys from S100A4-FGFR2−/− mice. a Representative immunohistochemical staining images showing Ki-67-positive cells in the kidneys from S100A4- FGFR2−/− mice and control littermates. b Graph showing quantitative results for Ki-67-positive cells in the kidneys. C, cortical area; M, medullary area; T, tubule; I, interstitium. Data are presented as number of Ki-67-positive cells per HPF (×400). * p < 0.05 versus control littermates, n = 4. FGFR, fibroblast growth factor receptor; HPF, high-power field; IRI, ischemia/reperfusion injury.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Interstitial cell apoptosis was less in mice with fibroblast-specific ablation of FGFR2 after IRI. a Representative micrographs showing TUNEL staining for apoptotic cells in the kidneys from S100A4-FGFR2−/− mice and control littermates. Arrows indicate apoptotic cells in the kidneys. b Graph showing the quantitative results for TUNEL-positive cells in the kidneys. C, cortical area; M, medullary area; T, tubule; I, interstitium. Data are presented as TUNEL-positive cell number per HPF (×400). * p < 0.05 versus control littermates, n = 4. FGFR, fibroblast growth factor receptor; HPF, high-power field; IRI, ischemia/reperfusion injury.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: TUNEL Assay, Staining, Control

Journal: Kidney Diseases

Article Title: Ablation of FGFR2 in Fibroblasts Ameliorates Kidney Fibrosis after Ischemia/Reperfusion Injury in Mice

doi: 10.1159/000484604

Figure Lengend Snippet: Erk1/2 phosphorylation was less in the kidneys from S100A4-FGFR2−/− mice after IRI. a, b Western blot analysis demonstrating the reduction in Erk1/2 phosphorylation in S100A4-FGFR2−/− kidneys at 14 days (a) and 28 days (b) after IRI compared to their control littermates. Numbers indicate individual animals. The gels were run under the same experimental conditions. The graphs show the semiquantitative results for p-Erk1/2 in the kidney lysates from S100A4-FGFR2−/− mice and control littermates at 14 and 28 days after IRI. * p < 0.05 versus control littermates, n = 4. c Representative immunohistochemical staining images showing the colocalization of α-SMA and p-Erk1/2 protein in fibrotic kidneys at 28 days after IRI. Asterisks indicate cells in the kidneys costaining positive for α-SMA and p-Erk1/2. α-SMA, alpha smooth muscle actin; Erk1/2, extracellular regulated protein kinase 1/2; FGFR, fibroblast growth factor receptor; IRI, ischemia/reperfusion injury.

Article Snippet: In some experiments, tissue sections were immunostained with primary antibodies against anti-FGFR2 (Cat: sc-122; Santa Cruz Biotechnology).

Techniques: Phospho-proteomics, Western Blot, Control, Immunohistochemical staining, Staining

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: FGFR2 Promotes Gastric Cancer Progression by Inhibiting the Expression of Thrombospondin4 via PI3K-Akt-Mtor Pathway.

doi: 10.1159/000494590

Figure Lengend Snippet: Fig. 6. The FGFR2-TSP4 axis promotes GC progression. The FGFR2 signal activates the PI3K-AKT- mTOR pathway resulting in the suppression of 4EBP1 phosphorylation and decreased the release of eIF-4E, which may inhibit the mRNA translation of TSP4. The FGFR2-TSP4 axis plays an important role in GC cells proliferation, invasion and migration.

Article Snippet: Specific primary antibodies against FGFR2 (1:100, TA503137, OriGene, USA) and TSP4 (1:100, AP19723b, Abgent, San Diego,CA) were used for IHC with a two-step protocol.

Techniques: Phospho-proteomics, Migration

Journal: American Journal of Cancer Research

Article Title: miR-381-3p suppresses the proliferation of oral squamous cell carcinoma cells by directly targeting FGFR2

doi:

Figure Lengend Snippet: FGFR2 is a direct downstream target of miR-381-3p. A. A fragment of FGFR2 3’-UTR containing the wild-type (WT) miR-381-3p or mutant binding site was cloned downstream of the reporter gene vector. B. Levels of FGFR2 mRNA and protein were measured using qRT-PCR and western blot, respectively, in response to overexpression of miR-381-3p in SCC-9 and Tca-8113 cells. C. Luciferase activity of wild-type or mutant FGFR2 3’-UTR luciferase vectors in the presence of miR-381-3p. D. FGFR2 mRNA expression in 18 pairs of tongue OSCC specimens and matched adjacent normal specimens. E. Scatter plot showing the correlation between miR-381-3p and FGFR2 expression in OSCC tissues. Data represent means ± SD of three independent experiments. *P < 0.05.

Article Snippet: Following transfer, membranes were blocked with 5% skimmed milk and incubated with the appropriate primary antibodies against FGFR2 and GAPDH (Proteintech Group, Wuhan, China).

Techniques: Mutagenesis, Binding Assay, Clone Assay, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Over Expression, Luciferase, Activity Assay, Expressing

Journal: American Journal of Cancer Research

Article Title: miR-381-3p suppresses the proliferation of oral squamous cell carcinoma cells by directly targeting FGFR2

doi:

Figure Lengend Snippet: miR-381-3p suppresses OSCC growth by downregulating FGFR2. A. SCC-9 cells were transfected with FGFR2 siRNA or negative control. After 48 h, the FGFR2 protein level was examined via western blot analysis. B. MTT, cell cycle and apoptosis assays. C. SCC-9 cells were infected with NC, miR-381-3p lentivirus or miR-381-3p and FGFR2 lentivirus. After 48 h, FGFR2 protein levels were examined via western blot analysis. D. MTT, cell cycle and cell apoptosis assays. Data represent means ± SD of three independent experiments. *P < 0.05 vs. NC group, $P < 0.05 vs. miR-381-3p group.

Article Snippet: Following transfer, membranes were blocked with 5% skimmed milk and incubated with the appropriate primary antibodies against FGFR2 and GAPDH (Proteintech Group, Wuhan, China).

Techniques: Transfection, Negative Control, Western Blot, Infection

Journal: American Journal of Cancer Research

Article Title: miR-381-3p suppresses the proliferation of oral squamous cell carcinoma cells by directly targeting FGFR2

doi:

Figure Lengend Snippet: Overexpression of miR-381-3p inhibits OSCC tumorigenesis in vivo. Tumors were detected every 7 days after implantation of SCC-9 cells infected with miR-381-3p or control lentivirus. A. Representative images of the tumors formed. B. Tumor volume. C. Mean weight. D. Ki-67-stained sections of transplanted tumors (×400). E. qRT-PCR analysis of expression of miR-381-3p and FGFR2 in tumor tissues. F. Western blot analysis of FGFR2 expression in tumors. Data represent means ± SD of three independent experiments. *P < 0.05.

Article Snippet: Following transfer, membranes were blocked with 5% skimmed milk and incubated with the appropriate primary antibodies against FGFR2 and GAPDH (Proteintech Group, Wuhan, China).

Techniques: Over Expression, In Vivo, Infection, Control, Staining, Quantitative RT-PCR, Expressing, Western Blot