Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: Primers sequences used for qRT-PCR.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques:

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: CircARVCF regulated FGFR1 expression by targeting miR-1205. (A) The complementary sequences between FGFR1 and miR-1205 were exhibited. (B,C) The relationship between FGFR1 and miR-1205 was analyzed by dual-luciferase reporter assay. (D,E) The mRNA and protein levels of FGFR1 in GC tissues and normal tissues were measured by qRT-PCR or western blot assay. (F) The protein level of FGFR1 in GES-1, MKN-45 and AGS cells was measured via western blot assay. (G,H) The mRNA and protein levels of FGFR1 in DDP-resistant and DDP-sensitive GC tissues were quantified by qRT-PCR or western blot. (I) The protein level of FGFR1 in MKN-45, MKN-45/DDP, AGS and AGS/DDP cells was measured via western blot assay. (J) The protein level of FGFR1 in MKN-45/DDP and AGS/DDP cells transfected with miR-NC, miR-1205, miR-1205 + pcDNA or miR-1205 + FGFR1 was measured via western blot assay. (K) The protein level of FGFR1 in MKN-45/DDP and AGS/DDP cells transfected with si-NC, si-circARVCF#1, si-circARVCF#1+in-miR-NC or si-circARVCF#1+in-miE-1205 was measured through western blot assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

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

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: MiR-1205 regulated the DDP resistance and malignant behaviors of DDP-resistant GC cells by targeting FGFR1. MKN-45/DDP and AGS/DDP cells were transfected with miR-NC, miR-1205, miR-1205 + pcDNA or miR-1205 + FGFR1. (A,B) DDP resistance was analyzed by CCK-8 assay. (C–F) The colony formation, migration, invasion and apoptosis of MKN-45/DDP and AGS/DDP cells were evaluated by colony formation assay, transwell assay and flow cytometry analysis, respectively. (G,H) The protein levels of MRP1, MDR1, Bcl-2 and Bax in MKN-45/DDP and AGS/DDP cells were measured via western blot assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques: Transfection, CCK-8 Assay, Migration, Colony Assay, Transwell Assay, Flow Cytometry, Western Blot

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: CircARVCF enhanced DDP resistance in GC in vivo . (A,B) Tumor volume and tumor weight were examined. (C,D) The levels of circARVCF and miR-1205 in the tumors were examined by qRT-PCR. (E,F) The level of FGFR1 in the tumors was examined by western blot assay and IHC assay. * p < 0.05.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques: In Vivo, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Genetics

Article Title: CircARVCF Contributes to Cisplatin Resistance in Gastric Cancer by Altering miR-1205 and FGFR1

doi: 10.3389/fgene.2021.767590

Figure Lengend Snippet: The frame diagram of circARVCF/miR-1205/FGFR1 axis in regulating DDP resistance and cell progression in GC.

Article Snippet: Next, the slides were incubated with anti-FGFR1 (bs-0230R; Bioss) overnight and secondary antibody (bs-0293G-HRP) for 1 h. After that, the signal was developed with DAB solution and counterstained with hematoxylin (Sigma-Aldrich) as previously described ( ).

Techniques:

Journal: Cancer Science

Article Title: FGFR gene alterations in lung squamous cell carcinoma are potential targets for the multikinase inhibitor nintedanib

doi: 10.1111/cas.13071

Figure Lengend Snippet: Effects of nintedanib therapy on molecular markers in vivo . Tumor samples were collected from mice 2 h after the final gavage following 15 days of treatment as indicated and subjected to molecular marker analysis. (a) Immunoblot analysis of the effects of nintedanib therapy on FGFR1, ERK , and AKT phosphorylation in the H520 and LK ‐2 xenograft tumor model. Cell lysates (25 μg soluble protein) were subjected to immunoblot analysis with antibodies to the indicated proteins. (b) Representative images of p‐ FGFR 1 immunostaining in each group. Stromal/vascular regions are denoted by the green dotted line. Scale bar = 100 μm. (c) Representative images of Ki‐67 immunostaining in each group. Scale bar = 100 μm. (d) Quantitative analyses of the number of Ki‐67‐positive cells in each group. (e) Representative images of CD 31 immunostaining in each group. Insets show higher‐magnification images of the blood vessel shown by the red frame. Scale bar = 100 μm (whole image) and 10 μm (inset image). (f) Quantitative analyses of vessel density, vessel area, vessel perimeter, and the ratio of vessel area/tumor area determined with CD 31‐positive cells. * P < 0.05 versus corresponding value for vehicle‐treated mice (Student's t ‐test).

Article Snippet: DNA copy numbers for FGFR1 , FGFR2 , FGFR3 , and FGFR4 were determined with the use of TaqMan Copy Number Assays (Applied Biosystems, Foster City, CA, USA) and primers Hs02164585_cn, Hs05208783_cn, Hs00113109_cn, and Hs01949336_cn, respectively.

Techniques: In Vivo, Marker, Western Blot, Phospho-proteomics, Immunostaining

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: Information about Assays for SNP Analyzed in this Study

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques: Marker

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: Linkage Disequilibrium between Markers Genotyped in the Study

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques: Marker

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: Association (FBAT) Results for Isolated Tooth Agenesis

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques: Isolation

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: IRF6 V274I and FGFR1 rs881301 Results (FBAT) for Subpopulations of Tooth Agenesis Cases

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques:

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: Case-Control Analysis of the Ohio Tooth Agenesis Population

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques:

Journal:

Article Title: Interferon Regulatory Factor 6 ( IRF6 ) and Fibroblast Growth Factor Receptor 1 ( FGFR1 ) Contribute to Human Tooth Agenesis

doi: 10.1002/ajmg.a.31620

Figure Lengend Snippet: IRF6 - and FGFR1 -Other Candidate Genes for Tooth Agenesis Interactions

Article Snippet: rs10958704 , 2 kb 5′ of FGFR1 , TaqMan OD , C_2080139_10.

Techniques:

Journal: Theranostics

Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

doi: 10.7150/thno.72269

Figure Lengend Snippet: High pY levels might affect the proliferation, migration and differentiation of lung fibroblasts. A , Changes in cell proliferation between hnLFs and hfLFs determined by CCK-8 kit (n = 7). *** P < 0.001. B-C , Assessing cell proliferation by EdU assay between hnLFs and hfLFs (n = 6). *** P < 0.001. D-G , Comparison of migration between hnLFs and hfLFs (n = 6). *** P < 0.001. H , Representative images of hnLFs and hfLFs under TEM. Arrows pointed to the lipid droplets in cytoplasm. I , Representative fluorescence images of Nile Red in hnLFs and hfLFs. J-K , The contractility of hnLFs and hfLFs in 3D collagen matrices (n = 6). *** P < 0.001. L , Phospho-RTK array analysis with 200 μg of lysates from hnLFs and hfLFs, which were serum-starved for 6 h. M , Quantification of phospho-RTK levels (average of two drops fold average of positive control). N-O , Western blot and quantification of p-VEGR2, p-PDGFRβ, p-EGFR and p-FGFR1 in hnLFs and hfLFs (n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001. P-Q , Western blot and quantification of p-PLCγ1, p-GAB1, p-SHC and p-SRC in hnLFs and hfLFs (n = 6). ** P < 0.01, *** P < 0.001. R-S , Western blot and quantification of p-AKT, p-ERK1/2 and p-STAT3 in hnLFs and hfLFs (n = 6). ** P < 0.01, *** P < 0.001.

Article Snippet: Antibodies against FN (Cat#15613-1-AP), COL1A1 (Cat#67288-1-lg), α-SMA (Cat#55135-1-AP, Cat#67735-1-AP), FGFR1 (Cat#60325-1-lg), CD31 (Cat#60287-1-lg), CD45 (Cat#11265-1-lg), EpCAM (Cat#60287-1-lg), Vimentin (Cat#10366-1-AP) and GAPDH (Cat#60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Migration, CCK-8 Assay, EdU Assay, Comparison, Fluorescence, Positive Control, Western Blot

Journal: Theranostics

Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

doi: 10.7150/thno.72269

Figure Lengend Snippet: SH2 superbinder blocked multiple fibrosis associated pathways through interrupting pY-SH2 combination in pY mediated signal transmission. A-B , Phospho-RTK array analysis with 200 μg of lysates from hfLFs treated with 1 μM GST-SH2 WT or GST-SH2 TrM for 24 h. C , Western blot of p-VEGR2, p-PDGFRβ, p-EGFR and p-FGFR1 in hnLFs and hfLFs after GST, GST-SH2 WT and GST-SH2 TrM incubation. D , Western blot of p-PLCγ1, p-GAB1, p-SHC and p-SRC in hnLFs and hfLFs after GST, GST-SH2 WT or GST-SH2 TrM incubation. E , Western blot of p-AKT, p-ERK1/2 and p-STAT3 in hnLFs and hfLFs after GST, GST-SH2 WT or GST-SH2 TrM incubation. F , hnLFs and hfLFs were incubated with 1 μM GST, GST-SH2 WT or GST-SH2 TrM for 24 h. The immunoprecipitation of EGFR and SHC was detected. G , Representative fluorescence images of EGFR and SHC colocalization in hnLFs and hfLFs after GST, GST-SH2 WT or GST-SH2 TrM treatment. H , Immunoprecipitations of GRB2 with GAB1, SHC, EGFR and PDGFRβ. I , GST pull down assay showed the binding capacity of SH2 superbinder with pY in hnLFs and hfLFs. J , Representative fluorescence images of GST tag and pY colocalization in hnLFs and hfLFs after incubation with GST, GST-SH2 WT or GST-SH2 TrM. K-L , GST pull down assay showed the binding capacity of SH2 superbinder with RTKs (such as VEGFR2, PDGFRβ, EGFR and FGFR1) and adaptor proteins (such as GAB1, SHC and SRC) in hfLFs and hfLFs.

Article Snippet: Antibodies against FN (Cat#15613-1-AP), COL1A1 (Cat#67288-1-lg), α-SMA (Cat#55135-1-AP, Cat#67735-1-AP), FGFR1 (Cat#60325-1-lg), CD31 (Cat#60287-1-lg), CD45 (Cat#11265-1-lg), EpCAM (Cat#60287-1-lg), Vimentin (Cat#10366-1-AP) and GAPDH (Cat#60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Transmission Assay, Western Blot, Incubation, Immunoprecipitation, Fluorescence, Pull Down Assay, Binding Assay

Journal: Scientific Reports

Article Title: Klotho Inhibits Interleukin-8 Secretion from Cystic Fibrosis Airway Epithelia

doi: 10.1038/s41598-017-14811-0

Figure Lengend Snippet: TGF-β induced upregulation of FGFR1 in CF-HBECs. ( a ) FGFR1 and FGFR4 mRNA levels in HBECs from nonsmokers and CF patients at baseline and ( b ) after stimulation with TGF-β (10 ng/mL) for 24 hours. ( c ) Immunohistochemistry staining with anti-FGFR1-HRP and hematoxylin eosin counterstain of whole lung sections from one control patient (non-smoker) compared to a CF patient (20X, scale bars are 40 μm): staining of the bronchial epithelium (arrows) was detected and increased in the CF lung section (representative specimen from a total of 4 lungs in each group). ( d ) FGFR1 mRNA levels of CF-HBECs, exposed to TGF-β for 24 hours (10 ng/mL) ± LY2157299 (10 μM) or SB203580 (10 μM). ( e ) Representative immunoblots are shown after stimulation of CF-HBEC with TGF-β (30–90 min, 10 ng/ml): increased Smad 3 and increased ERK phosphorylation were seen without changes in PLCγ phosphorylation. ( f ) Bar graphs showing densitometric analysis of TGF-β induced fold changes in phospho ERK/total ERK and phospho Smad 3/total Smad 3 ratios in control HBECs (n = 5) compared to CF-HBEC (n = 6). ( g ) Dot plot indicating densitometric changes in phospho Smad/ total Smad 3 ratios in HBECs and CF-HBECs when untreated and TGF-β treated, shown as fold change increase using unpaired t-test and Mann-Whitney post test. Data is presented as means ± S.E. with *P < 0.05, **P < 0.01, ***P < 0.001 and ## P < 0.01 compared to TGF-β treatment. The full-length blots are presented in Supplementary Figure .

Article Snippet: Real-time quantitative PCR was performed using the following TaqMan probes: Hs00241111_m1 for FGFR1, Hs01106910_g1 for FGFR4, Hs00934627_m1 for KL, Hs00174103_m1 for IL-8, Mm00446190_m1 for IL-6, and Hs02758991_g1 for GAPDH.

Techniques: Immunohistochemistry, Staining, Control, Western Blot, Phospho-proteomics, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Klotho Inhibits Interleukin-8 Secretion from Cystic Fibrosis Airway Epithelia

doi: 10.1038/s41598-017-14811-0

Figure Lengend Snippet: Exposure to TGF-β ± FGF23 increases IL-8 secretion in CF-HBECs. ( a ) Bar graphs indicating relative transcript numbers of IL-8 mRNA, compared to GAPDH, of both HBECs and CF-HBECs after stimulation with FGF23 (25 ng/mL) ± TGF-β (10 ng/mL) for 24 h. ( b / c ) Inhibition of either FGF23 (via FGFR1) or TGF-β (via p38) signaling prevented the increase in ( b ) IL-8 mRNA expression and ( c ) IL-8 secretion, suggesting that these pathways are both involved. TGF-β signaling was inhibited by LY2157299 (10 μM), p38 MAPK by SB203580 (10 μM) and FGFR1 signaling by AZD4745 (5 nM). All n = 3 independent experiments from 3 different lungs. In graphs, means ± S.E are shown. *P < 0.05, **P < 0.01, ***P < 0.005 for comparisons with the control and ### P < 0.005 for comparison with the treated group.

Article Snippet: Real-time quantitative PCR was performed using the following TaqMan probes: Hs00241111_m1 for FGFR1, Hs01106910_g1 for FGFR4, Hs00934627_m1 for KL, Hs00174103_m1 for IL-8, Mm00446190_m1 for IL-6, and Hs02758991_g1 for GAPDH.

Techniques: Inhibition, Expressing, Control, Comparison

Journal: Scientific Reports

Article Title: Klotho Inhibits Interleukin-8 Secretion from Cystic Fibrosis Airway Epithelia

doi: 10.1038/s41598-017-14811-0

Figure Lengend Snippet: Modification of KL levels changes IL-6 and IL-8 expression. ( a ) Relative transcript numbers of murine kl mRNA and ( b ) endogenous human KL mRNA are shown in fully differentiated HBECs, either control infected or infected with murine full length kl expressing lentiviruses. ( c ) Secreted IL-8 protein levels in both control and murine kl expressing HBECs ± treatment with TGF-β + FGF23 show attenuation of IL-8 secretion in kl infected HBECs. All n = 3 independent experiments from 3 different lungs. ( d ) Secreted IL-6 protein levels at baseline are significantly elevated in murine tracheal epithelial cells (MTECs) isolated from kl −/− and kl +/− mice when compared to wild type controls. Bar graphs indicating ( e ) IL-6 mRNA and ( f ) FGFR1 mRNA from wild type versus kl +/− MTECs after stimulation with TGF-β ± FGF23. MTECs were pooled from 3–4 mouse lungs and 3 independent experiments were performed. All bar graphs are mean ± S.E. *P < 0.05, **P < 0.01 and ***P < 0.005 compared to control.

Article Snippet: Real-time quantitative PCR was performed using the following TaqMan probes: Hs00241111_m1 for FGFR1, Hs01106910_g1 for FGFR4, Hs00934627_m1 for KL, Hs00174103_m1 for IL-8, Mm00446190_m1 for IL-6, and Hs02758991_g1 for GAPDH.

Techniques: Modification, Expressing, Control, Infection, Isolation

Journal: Scientific Reports

Article Title: Klotho Inhibits Interleukin-8 Secretion from Cystic Fibrosis Airway Epithelia

doi: 10.1038/s41598-017-14811-0

Figure Lengend Snippet: Diagram illustrating the effect of TGF-β and FGF23 on CF bronchial epithelial cells. TGF-β upregulates FGFR1 and together with FGF23, they induce IL-8 expression via activation of ERK and TGF-βR signaling. In addition, TGF-β induces KL secretion, which attenuates IL-8 secretion by inhibition of TGF-βR signaling. Definition of abbreviations: IL = interleukin, FGF = fibroblast growth factor, FGFR = FGF receptor, ERK = Extracellular Signal regulated kinases.

Article Snippet: Real-time quantitative PCR was performed using the following TaqMan probes: Hs00241111_m1 for FGFR1, Hs01106910_g1 for FGFR4, Hs00934627_m1 for KL, Hs00174103_m1 for IL-8, Mm00446190_m1 for IL-6, and Hs02758991_g1 for GAPDH.

Techniques: Expressing, Activation Assay, Inhibition