Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: FGFR2-IIIb Immunoreactivity in HCC Tissue of 85 Patients in Relation to Clinicopathological Characteristics and Proliferation Rate

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Biomarker Discovery

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Sets of Primers Used for Quantitative PCR Analysis

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Real-time Polymerase Chain Reaction

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: FGFR2-IIIb expression in HCC. FGFR2-IIIb mRNA expression was determined by using quantitative RT-PCR in (A) PHH and three HCC cell lines, two murine HCC models, (B) c-myc and (C) IgEGF transgenic animals, and (D) 15 human HCC samples and corresponding nontumorous liver tissue samples. E: Immunohistochemical staining for FGFR2-IIIb (left) and H&E staining (right) of HCC and neighboring nontumorous (nt) liver tissue are shown. F: Proliferation rate (analyzed by immunohistochemical staining applying Ki-67 antibodies) in HCC tissues with different FGFR2-IIIb immunoreactivity is shown.

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Expressing, Quantitative RT-PCR, Transgenic Assay, Immunohistochemical staining, Staining

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Analysis of FGFR2-IIIc expression and FGFR2-IIIb promotor methylation in HCC. RT-PCR analysis of FGFR2-IIIc and FGFR2-IIIb mRNA expression in primary human hepatocytes and three different HCC cell lines (A) and primary human HCC tissues of six different donors (B) is shown. β-actin served as a control. C: Methylation-sensitive PCR analysis of the CpG island in the 5′-region of the FGFR2 gene is shown. PCR of DNA samples were digested with AfII (A, cutting outside the region analyzed) as a positive control, AfII plus MspI (A + M, cutting within the region analyzed, methylation-insensitive) as a negative control, and AfII plus SmaI (A + S), HpaII (A + H), or EclVI (A + E) (all three cutting within the region analyzed and methylation-sensitive). D: Amplification of a 34-bp fragment containing exon 8 of the FGFR2 gene is shown.

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Expressing, Methylation, Reverse Transcription Polymerase Chain Reaction, Control, Positive Control, Negative Control, Amplification

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Re-expression of FGFR2-IIIb in HCC cells. Analysis of FGFR2-IIIb mRNA (A) and protein expression (B) in stable transfected HepG2 cell clones (c1–c3), mock transfected controls (mock), and nontransfected cells (wt) by quantitative RT-PCR and Western blotting is shown. C: Overall, protein synthesis was analyzed by determining [2H8]-valine protein incorporation.

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Expressing, Transfection, Clone Assay, Quantitative RT-PCR, Western Blot

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Functional analysis of FGFR2-IIIb re-expressing HCC cells in vitro. Proliferation (A), attachment independent growth (analyzed by soft agar colony formation assays, B), apoptosis (analyzed by fluorescence activated cell sorting (FACS) analysis, C), and migratory potential (assessed by Boyden Chamber assays, D) of HepG2 cell clones stably expressing FGFR2-IIIb (c1–3), mock transfected controls (mock), and nontransfected (wt) controls are shown. *P < 0.05.

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Functional Assay, Expressing, In Vitro, Fluorescence, FACS, Clone Assay, Stable Transfection, Transfection

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Effect of KGF and SU5402, an inhibitor of tyrosine kinase activity, on proliferation of FGFR2-IIIb re-expressing HCC cells. Proliferation was analyzed in HepG2 cell clones stably expressing FGFR2-IIIb (clone 2 and clone 3) pretreated or not with KGF (20 ng/ml, A) or FGFR tyrosine kinase inhibitor (SU5402; 30 μmol/L, B).

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Activity Assay, Expressing, Clone Assay, Stable Transfection

Journal: The American Journal of Pathology

Article Title: Reduced Expression of Fibroblast Growth Factor Receptor 2IIIb in Hepatocellular Carcinoma Induces a More Aggressive Growth

doi: 10.2353/ajpath.2010.090356

Figure Lengend Snippet: Tumorigenicity of FGFR2-IIIb re-expressing HCC cells in vivo. A: Tumor size 21 days after implantation of 1 × 106 HepG2 cells stably transfected with a FGFR2-IIIb-expression plasmid (c1–3) and mock transfected (mock) and nontransfected control cells into nude mice (n = 10/group). *P < 0.05 compared with both wt and mock. B: Analysis for apoptotic cells by using terminal deoxynucleotidyl transferase dUTP nick-end labeling staining. DAPI staining of the same section was applied to visualize cell nuclei.

Article Snippet: 23 table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Gene Forward primer Reverse primer Human β- Actin 5′-CTACGTCGCCCTGGACTTCGAGC-3′ 5′-GATGGAGCCGCCGATCCACACGG-3′ FGFR2-IIIb 5′-TGCTGGCTCTGTTCAATGTG-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ FGFR2-IIIc 5′-ACACCACGGACAAAGAGATT-3′ 5′-GGCGATTAAGAAGACCCCTA-3′ Mouse β- Actin 5′-AGGCCAACCGTGAAAAGAT-3′ 5′-GGCGTGAGGGAGAGCATA-3′ FGFR2-IIIb 5′-GCCCATCCTCCAAGCTG-3′ 5′-TGGTTGGCCTGCCCTATA-3′ Open in a separate window Sets of Primers Used for Quantitative PCR Analysis Protein Analysis Polyclonal antibodies against FGFR2-IIIb were prepared by immunization of rabbits against synthetic peptides in cooperation with Biogenes (Berlin, Germany).

Techniques: Expressing, In Vivo, Stable Transfection, Transfection, Plasmid Preparation, Control, TUNEL Assay, Staining

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: Frontiers in Immunology

Article Title: Disruption of Notch signaling by KGF induces a developmental pause in thymocytes

doi: 10.3389/fimmu.2025.1675823

Figure Lengend Snippet: KGF reduces expression of DLL4 and its upstream transcriptional regulator FOXN1 in both cTECs and mTECs. (A) Thymic stromal subsets were defined based on EpCAM, Ly51, and UEA-1 expression profiles among CD45 - cells. Representative dot plots from PBS- and KGF-treated thymi (three independent experiments) are shown (left). EpCAM + TECs were further subdivided into two major subsets: UEA-1 + /Ly51 − medullary TECs (mTECs) and UEA-1 − /Ly51 + cortical TECs (cTECs). DLL4 expression in cTECs and mTECs from PBS- (blue) and KGF-treated (red) wild type mice was evaluated by flow cytometry; representative histograms (middle) and mean fluorescence intensity (MFI) values (right) are shown. Isotype staining is shown in black. (B) Dll4 mRNA levels were measured in total TECs from PBS- (blue) and KGF-treated (red) mice. Data from three independent experiments are presented. (C) Cell surface expression of Notch1 receptor in DN2a-DN3a early thymocyte subsets from PBS- (blue) and KGF-treated (red) mice was evaluated by flow cytometry. Isotype staining is shown in black. Representative histograms from four independent experiments are shown (left), and MFI for each subset are presented (right). (D) Notch1 mRNA levels in DN2a-DN3a thymocyte subsets from PBS- (blue) and KGF-treated (red) mice are shown. (E) Intracellular FOXN1 expression in TECs from PBS- (blue) and KGF-treated (red) mice was assessed by flow cytometry. Representative histograms are shown, including an unstained control (black line) and IgG2b isotype control (green line) (left). Quantification of FOXN1 expression in total TECs by MFI is based on three independent experiments. (F) Foxn1 mRNA levels in sorted TEC subsets from PBS- (blue) and KGF- treated (red) mice were also evaluated and quantitative results are shown. Data are presented as mean ± SD. Error bars indicate standard deviation. Statistical significance between two groups was determined using unpaired two-tailed t-tests. One-way ANOVA was applied for comparison of 3 and more groups and p-values were determined by Tukey’s multiple comparisons test: *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001; ns = not significant.

Article Snippet: After blocking, tissue sections were incubated overnight at 4°C with a combination of primary antibodies: anti-mouse FGFR2IIIb rat IgG2a (clone 133730, #MAB7161, R&D Systems) together with either anti-mouse Keratin 5 rabbit IgG (clone AF138, #PRB-160P, Covance) or anti-mouse Keratin 8 rabbit IgG (clone EP1628Y, #ab53280, Abcam).

Techniques: Expressing, Flow Cytometry, Fluorescence, Staining, Control, Standard Deviation, Two Tailed Test, Comparison

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: ( A ) FGFR2 copy number obtained in real-time PCR assay; ( B ) mRNA expression by qRT-PCR assay; ( C ) FGFR2 expression by western blot analysis; ( D ) Distributions of FGFR2 copy number ( N = 140). For FGFR2 amplification, the copy number gain must exceed 3.0 copies. FGFR2 amplification was observed in 21 cases (21/140 = 15%); ( E ) FGFR2 was not expressed in normal glandular epithelium; ( F ) ( a, b ) Cases with absent or faint FGFR2 staining were assessed as FGFR2 IHC-negative; ( c, d ) cases with moderate or strong FGFR2 staining were FGFR2 IHC-positive; ( G ) Association between FGFR2 IHC positivity and FGFR2 amplification; ( H ) Cancer-specific survival in positive and negative FGFR2 IHC cases; ( I ) Overall survival in positive and negative FGFR2 IHC cases.

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Quantitative RT-PCR, Western Blot, Amplification, Staining

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: Associations between FGFR2 positivity and clinicopathological factors in EGJ adenocarcinoma patients with tumor resection

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Amplification, Paraffin-embedded Immunohistochemistry

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: Red and green signals indicate FGFR2 gene and centromere of chromosome 10 probes, respectively. ( A ) FGFR2-amplified case obtained by real-time PCR reaction; ( B ) FGFR2-non-amplified case obtained by real-time PCR reaction.

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Amplification, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: Univariate and multivariate logistic analysis of FGFR2 IHC status in EGJ adenocarcinoma patients with tumor resection

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: ( A ) De-phosphorylation of AKT and ERK after FGFR2 knockdown by siRNAs targeting FGFR2 (si-FGFR2); ( B ) Cell proliferation after transfection with si-control or si-FGFR2s; ( C ) Distributions of cell cycle populations; ( D ) Proportions (%) of G0/G1, S, and G2/M cells in the cell cycle distribution; ( E ) Distributions of apoptotic cells; ( F ) Apoptotic cells, identified as positive for Annexin V and negative for propidium iodide (PI); ( G ) Proportions (%) of sub-G1 cells in the cell cycle distribution. Panels (D), (F) and (G) show the results at 72 h after transfection with si-control or si-FGFR2s. * P < 0.05.

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: De-Phosphorylation Assay, Knockdown, Transfection, Control

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: ( A ) FLO-1 cells stably transfected with FGFR2 were confirmed by GFP expression; ( B ) FGFR2 overexpression was confirmed by western blot analysis; ( C ) Cell proliferation of FGFR2 overexpressing cells; ( D ) Distributions of cell cycle populations; ( E ) Proportions (%) of G0/G1, S, and G2/M in the cell cycle distribution; ( F ) Distributions of apoptotic cells; ( G ) Apoptotic cells, identified as positive for Annexin V and negative for propidium iodide (PI); ( H ) Proportions (%) of sub-G1 cells in the cell cycle distribution. * P < 0.05.

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Stable Transfection, Transfection, Expressing, Over Expression, Western Blot

Journal: Oncotarget

Article Title: Fibroblast growth factor receptor 2 expression, but not its genetic amplification, is associated with tumor growth and worse survival in esophagogastric junction adenocarcinoma

doi: 10.18632/oncotarget.7782

Figure Lengend Snippet: ( A ) Proliferation assay of tumor cells exposed to AZD4547 with and without FGF7 stimulation; ( B ) De-phosphorylation of AKT and ERK by AZD4547, with and without FGF7 stimulation; ( C ) Proliferation assay using siRNAs targeted against FGFR2 (si-FGFR2), with and without FGF7 stimulation; ( D ) De-phosphorylation of AKT and ERK after FGFR2 knockdown by si-FGFR2, with and without FGF7 stimulation. * P < 0.05.

Article Snippet: The primary antibodies, FGFR2 (#11835s), AKT (#9272), phospho-AKT Ser473 (#9271), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#4376) and Β-actin (#4967s), were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Proliferation Assay, De-Phosphorylation Assay, Knockdown

Journal: Oncogene

Article Title: Genome-wide CRISPR screen identifies LGALS2 as an oxidative stress-responsive gene with an inhibitory function on colon tumor growth

doi: 10.1038/s41388-020-01523-5

Figure Lengend Snippet: A Western blotting showing the expression of p-STAT3 and STAT3 in healthy colon or colon tumors. B Densitometry quantification of p-STAT3/total STAT3 for colons or colorectal tumors. *** p < 0.001 (one-way ANOVA). C Representative images of p-STAT3 immunostaining of colons or colon tumors. Scale bar: 50 μm. D Quantification of p-STAT3 + cells in colon tumors of WT and Gal2-KO mice. *** p < 0.001 ( t -test).

Article Snippet: Primary antibodies including the goat polyclonal anti-Gal2 (STJ24400, 1:500, St John’s Laboratory Ltd, London, UK), anti-STAT3 (#9139, 1:1000), anti-p-STAT3 (#9145, 1:1000), and anti-GAPDH (MAB374, 1:2000) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, Expressing, Immunostaining

Journal: Oncogene

Article Title: Genome-wide CRISPR screen identifies LGALS2 as an oxidative stress-responsive gene with an inhibitory function on colon tumor growth

doi: 10.1038/s41388-020-01523-5

Figure Lengend Snippet: A Western blotting showing the expression of Gal2 in stable Gal2-overexpressing HCT116 cells. The red arrow indicates the un-cleaved GFP-2A-Gal2 fusion protein. B The CCK8 proliferation assay of HCT116 with or without Gal2 overexpression. C The survival assay of HCT116 with or without Gal2 overexpression in the presence of 0.6 mM H 2 O 2 . ** p < 0.01, *** p < 0.001 ( t -test). D Western blotting showing the expression of p-STAT3 and STAT3 in HCT116 cells with or without Gal2 overexpression. E Densitometry quantification of p-STAT3/total STAT3 in HCT116 cells with or without Gal2 overexpression. ** p < 0.01 (one-way ANOVA).

Article Snippet: Primary antibodies including the goat polyclonal anti-Gal2 (STJ24400, 1:500, St John’s Laboratory Ltd, London, UK), anti-STAT3 (#9139, 1:1000), anti-p-STAT3 (#9145, 1:1000), and anti-GAPDH (MAB374, 1:2000) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, Expressing, Proliferation Assay, Over Expression, Clonogenic Cell Survival Assay

Journal: Cancer Pathogenesis and Therapy

Article Title: miR-338-3p acts as a tumor suppressor in lung squamous cell carcinoma by targeting FGFR2/FRS2

doi: 10.1016/j.cpt.2022.12.004

Figure Lengend Snippet: mRNA expression levels of FGFR2 and FRS2 in LUSC tissues and non-tumor lung tissues and their correlation with miR-338-3p. ( A) Comparison of FGFR2 and FRS2 mRNA expression levels between LUSC tissues and non-tumor lung tissues. (B) Pearson correlation analysis of the expression levels of miR-338-3p and FGFR2 in LUSC tissues ( P ​< ​0.001, r ​= ​−0.636). (C) The correlation between the expression levels of miR-338-3p and FRS2 in LUSC tissues was analyzed using the Pearson correlation analysis ( P ​< ​0.001, r ​= ​−0.691). (D) The expression of FGFR2 in adjacent normal lung tissue (IHC 100 ​× ​). (E) Expression of FGFR2 in LUSC tissues (IHC 400 ​× ​). (F) The expression of FRS2 in adjacent normal lung tissue (IHC 100 ​× ​). (G) FRS2 expression in LUSC tissues (IHC 400 ​× ​). IHC: Immunohistochemistry; LUSC: Lung squamous cell carcinoma.

Article Snippet: After the membranes were blocked with 5% non-fat milk for 2 h at room temperature, they were incubated at 4 °C with primary antibodies against FGFR2 (catalog #23328, Cell Signaling Technology), FRS2 (catalog: sc-17841, Santa Cruz Biotechnology), and β-actin (catalog: ab8245, Abcam), respectively, overnight.

Techniques: Expressing, Comparison, Immunohistochemistry

Journal: Cancer Pathogenesis and Therapy

Article Title: miR-338-3p acts as a tumor suppressor in lung squamous cell carcinoma by targeting FGFR2/FRS2

doi: 10.1016/j.cpt.2022.12.004

Figure Lengend Snippet: FGFR2 and FRS2 are direct target genes of miR-338-3p. (A) Predicted mRNA target regions and corresponding pairings of miR-338-3p. (B) Displays the relative fluorescence in H1703 ​cells when pGL3-FGFR2-3′-UTR or pGL3-FRS2-3′-UTR was transfected with the miR-338-3p mimic compared to the miRNA mimic control (negative control). Sulfatase activity decreased significantly ( P ​< ​0.001). (C) Compared with the miRNA mimic control (negative control), the mRNA expression levels of FGFR2 and FRS2 in H1703 ​cells were significantly downregulated after transfection with miR-338-3p mimic for 48 ​h ​( P ​< ​0.001). (D) Approximately 48 ​h after transfection with the miR-338-3p mimic and miRNA mimic control, the protein expression levels of FGFR2 and FRS2 in H1703 ​cells were detected using Western blot ( P ​< ​0.001).

Article Snippet: After the membranes were blocked with 5% non-fat milk for 2 h at room temperature, they were incubated at 4 °C with primary antibodies against FGFR2 (catalog #23328, Cell Signaling Technology), FRS2 (catalog: sc-17841, Santa Cruz Biotechnology), and β-actin (catalog: ab8245, Abcam), respectively, overnight.

Techniques: Fluorescence, Transfection, Control, Negative Control, Activity Assay, Expressing, Western Blot