Journal: bioRxiv

Article Title: Targeting Aberrant FGFR Signaling with Infigratinib Enhances the Efficacy of BTK/PI3K Inhibitors and Bendamustine in Lymphoma Cells

doi: 10.64898/2025.12.23.695986

Figure Lengend Snippet: A) Heatmap showing expression of FGFR1/3 and FGF2/9 across a panel of cell lines derived from MCL, ABC-DLBCL, and germinal center B-cell (GCB) DLBCL; heatmap is based on log2 CPM (counts per million). The scale ranges from low (-4, blue) to high (+4, red). Each horizontal represents the values for one cell line. B) Immunoblot for FGFR1 protein in a subset of selected cell lines and C) its protein quantification expressed as a ratio to SP53; data is the average with the second replicate shown in Supplementary Figure 1.

Article Snippet: The following primary antibodies were used in TBST 5% BSA buffer: FGFR1 mouse monoclonal (M19B2) (NB600-1287, Novus),Bek/FGFR2 mouse monoclonal (C-8) (sc-6930, Santa Cruz), FGFR3 mouse monoclonal (MAB766-100, ReD) and FGFR-4 mouse monoclonal (A-10) (sc-136988, Santa Cruz).

Techniques: Expressing, Derivative Assay, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Relative cell counts of viable Ba/F3 cells transduced with different vectors (empty vector, FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) normalized to parental Ba/F3 cells. Analyses were performed with 10 ng/mL IL-3 in culture medium and after withdrawal of IL-3 after 144 hours. Average numbers (±SD) of 6 independent experiments are shown. P values were calculated in pair-wise comparisons to Ba/F3 parental cells using a 1-sided Wilcoxon rank-sum test and adjusted for multiple testing using the Benjamini–Hochberg method. ( B ) Levels of total and phosphorylated proteins of the FGFR pathway in IL-3–dependent Ba/F3 cells (parental, empty vector, FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) and 3 IL-3–independent FGFR1 N546K - transduced Ba/F3 cell clones (blue). IL-3–independent FGFR1 N546K - transduced BA/F3 cells shown in different blue tones correspond to cells from 3 independent experiments labelled in light blue in ( A ). Antibodies against total and phosphorylated FGFR1, the adaptor protein FRS2, and the downstream targets AKT, ERK, and STAT3 were used. β-actin served as loading control.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Transduction, Plasmid Preparation, Clone Assay, Control

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Relative cell viability of IL-3–dependent Ba/F3 cells (parental, empty vector, FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) and 3 IL-3–independent FGFR1 N546K - transduced Ba/F3 cell clones (blue) after treatment with various concentrations (conc.) of futibatinib (in μM: 0.0001, 0.001, 0.01, 0.0398, 0.0631, 0.1, 1, 10, and 100) and DMSO as control for 72 hours. Mean cell viabilities ± SD of 3 ( FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) or 4 (parental, empty vector, FGFR1 N546K IL-3–independent cells) independent experiments conducted in triplicate each are plotted. ( B ) IC 50 of futibatinib in IL-3–dependent Ba/F3 cells and 3 IL-3–independent FGFR1 N546K -transduced Ba/F3 cell clones, derived from 3 ( FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) or 4 (parental, empty vector, FGFR1 N546K IL-3–independent cells) independent experiments consisting of triplicates each. P values were calculated in pair-wise comparisons to Ba/F3 parental cells using a 1-sided Wilcoxon rank-sum test and adjusted for multiple testing using the Benjamini–Hochberg method. ( C ) Levels of total and phosphorylated proteins of the FGFR pathway in IL-3–dependent Ba/F3 cells (parental, empty vector, FGFR1 WT, FGFR1 D623A , FGFR1 D623A;N546K , FGFR1 N546K ) and 3 IL-3–independent FGFR1 N546K transduced Ba/F3 cell clones (blue) after treatment with DMSO or 10, 50, or 100 nM futibatinib (experiment was conducted 3 times).

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Plasmid Preparation, Clone Assay, Control, Derivative Assay

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Images of abdominal cavities of R26-LSL-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt mice sacrificed at the ages of 7 and 21 days, showing tumors between the kidneys (top). Arrows point to the tumor boundaries. Kidneys (K) and tumors (indicated by asterisks) were prepared for better visualization of the tumors (bottom). ( B ) H&E and PHOX2B and Ki67 immunohistochemical staining of tumor sections obtained from R26-LSL-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt mice at the ages of 7 (left) and 14 (right) days; scale bar: 50 μm. ( C and D ) OS ( C ) and neuroblastoma-specific (NB-specific) survival ( D ) of R26-LSL-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt (blue), Th-MYCN tg/wt (red), and Th-ALK F1174L/wt (yellow) mice. Although OS was reduced in FGFR1 N546K transgenic mice in comparison with the other groups, due to development of papillomas and sarcomas ( FGFR1 N546K vs. MYCN tg , P < 0.001; FGFR1 N546K vs. ALK F1174L , P < 0.001; MYCN tg vs. ALK F1174L , P = 0.106), neuroblastoma-specific survival of FGFR1 N546K transgenic mice was worse only in comparison with ALK F1174L mice ( P = 0.049), but did not differ from that of MYCN tg mice ( P = 0.226). Survival of ALK F1174L compared with MYCN tg mice was not significant ( P = 0.394). Survival curves were estimated according to Kaplan-Meier test and compared with a log-rank test.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Immunohistochemical staining, Staining, Transgenic Assay, Comparison

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Axial T2-weighted MRI scans of an R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mouse that developed neuroblastomas at the adrenal glands (left) and the pelvic sympathetic trunk (right). ( B ) OS of R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt , Th- ALK F1174L/wt ; Th-MYCN tg/wt , and R26-LSL-FGFR1 N546K/wt ;Th-MYCN tg/wt mice. Survival of all groups differed significantly from each other in pairwise comparisons ( P < 0.001 each). Survival curves were estimated according to Kaplan-Meier tests and compared with log-rank tests. ( C ) H&E (top) and PHOX2B immunohistochemical (bottom) staining of an adrenal tumor obtained from an R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mouse at the age of 21 days, and of an adrenal tumor obtained from a Th-ALK F1174L/wt ;Th-MYCN tg/wt mouse at the age of 49 days; scale bar: 50 μm. ( D ) FGFR1 transcript levels in tumors obtained from R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mice and from Th-ALK F1174L/wt ;Th-MYCN tg/wt mice ( n = 11 each), as determined by RNA-Seq. Box plots show the median, first, and third quartiles; the whiskers represent the minimum and maximum values within ±1.5 times the interquartile range. Comparisons between groups were performed using a 2-tailed Wilcoxon rank-sum test.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Immunohistochemical staining, Staining, RNA Sequencing

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A – C ) Expression levels of genes associated with proliferation ( Mki67 , Ccnb1 , Ccnd1 , Top2a , and Pcna ( A ); anti-apoptotic genes ( Bcl2 and Bcl2l1 ) ( B ); and pro-apoptotic genes ( Casp3, Bax , and Bid ) ( C ) in tumors of R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt and Th-ALK F1174L/wt ;Th-MYCN tg/wt mice, as determined by RNA-Seq. Box plots show the median, first, and third quartiles; the whiskers represent the minimum and maximum values within ±1.5 times the interquartile range. Comparisons between groups were performed using a 2-tailed Wilcoxon rank-sum test. ( D ) H&E and PHOX2B, Ki67, and cleaved caspase 3 (CC3) immunohistochemical staining of tumor sections obtained from a Th-ALK F1174L/wt ;Th-MYCN tg/wt mouse (56 days old; top) and from an R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mouse (20 days old; bottom); scale bar, 50 μm. ( E ) BCL2 immunohistochemical staining of tumor sections obtained from of Th-ALK F1174L/wt ;Th-MYCN tg/wt (top) and R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt (bottom) mice; scale bar: 100 μm.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Expressing, RNA Sequencing, Immunohistochemical staining, Staining

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Temporal changes of relative tumor diameters in R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mice treated with 5 mg/kg futibatinib (light green), 30 mg/kg futibatinib (dark green), or control substance (0.5% CMC-Na; blue). Growth curves of individual tumors are shown on the left; mean and range of the relative tumor diameter changes are shown on the right. ( B ) OS of R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mice treated with 5 mg/kg, or 30 mg/kg futibatinib, or control. Survival of mice treated with 5 mg/kg futibatinib was significantly longer than that of control mice ( P < 0.001), whereas survival of mice treated with 30 mg/kg futibatinib was not significantly prolonged, due to toxicity ( P = 0.47). ( C ) Axial T2-weighted MRI scans of an R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mouse treated with 5 mg/kg futibatinib at the indicated ages. ( D ) H&E and PHOX2B, Ki67, and CC3 immunohistochemical staining of tumor sections obtained from R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mice treated with control (top) or 5 mg/kg futibatinib (bottom); scale bar, 50 μm. ( E ) Absolute volumes of tumors obtained from R26-FGFR1 N546K/wt ;Th-IRES-Cre tg/wt ;Th-MYCN tg/wt mice, subcutaneously reimplanted into NSG mice and treated with 10 mg/kg futibatinib ( n = 11) or control ( n = 9) after reaching a volume of 0.08–0.2 cm 3 . Growth curves of individual tumors are shown on the left; mean and range of tumor volumes are shown on the right.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Control, Immunohistochemical staining, Staining

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Absolute volumes of individual tumors (left) and mean and range of tumor volumes (right) of an FGFR N546K -mutant, patient-derived xenograft mouse model, treated with control (10% DMSO + 90% [20%] Captisol in 0.9% NaCl); n = 13) or 20 mg/kg futibatinib (futibatinib diluted in 10% DMSO + 90% [20%] Captisol in 0.9% NaCl); n = 13). ( B ) OS of mice bearing an FGFR N546K -mutant, patient-derived xenograft, treated with control or 20 mg/kg futibatinib. Survival curves were estimated according to Kaplan-Meier test and compared with a log-rank test. ( C ) H&E and PHOX2B and Ki67 immunohistochemical staining of tumor sections obtained from patient-derived xenograft tumors treated with a control substance (top) of 20 mg/kg futibatinib (bottom); scale bar, 100 μm.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Mutagenesis, Derivative Assay, Control, Immunohistochemical staining, Staining

Journal: The Journal of Clinical Investigation

Article Title: Mutated FGFR1 is an oncogenic driver and therapeutic target in high-risk neuroblastoma

doi: 10.1172/JCI189152

Figure Lengend Snippet: ( A ) Schematic timeline of the clinical course of a patient with FGFR1 N546K -mutated neuroblastoma that progressed under multiple lines of treatment and therefore was treated with a combination of futibatinib, cyclophosphamide, and topotecan. ( B ) Computed tomography (CT) scans of the tumor region in patient shown in ( D ) on day 15 after start of futibatinib/cyclophosphamide/topotecan (day 380 after diagnosis) and on day 87 (day 442 after diagnosis), demonstrating a partial regression of the tumor. ( C ) Absolute volumes of individual tumors (left) and mean and range of tumor volumes (right) of the patient-derived xenograft mouse model, treated with cyclophosphamide/topotecan alone (blue) or in combination with 20 mg/kg futibatinib (green). ( D ) OS of mice bearing the patient-derived xenograft, treated with cyclophosphamide/topotecan alone (blue) or in combination with 20 mg/kg futibatinib (green). Survival curves were estimated according to Kaplan-Meier test and compared with a log-rank test. MIBG, metaiodobenzylguanidine.

Article Snippet: The R26-LSL-FGFR1 N546K genetically engineered mouse line was generated by Taconic Biosciences and crossbred with the established Th-MYCN and Th-IRES-Cre mouse lines ( , ).

Techniques: Computed Tomography, Biomarker Discovery, Derivative Assay

Journal: Molecular Medicine

Article Title: The cytotoxic conjugate of highly internalizing tetravalent antibody for targeting FGFR1-overproducing cancer cells

doi: 10.1186/s10020-021-00306-2

Figure Lengend Snippet: Conjugation of tetravalent engineered antibody (T-Fc) with a cytotoxic payload. a Hypothetical model of the effect of FGFR1 clustering on the receptor endocytosis. FGFR1 dimerization via FGF1 binding induces receptor activation and clathrin-mediated endocytosis. Clustering of FGFR1 into large structures on the plasma membrane with tetravalent T-Fc largely improves the cellular uptake of FGFR1-antibody complexes. Furthermore, FGFR1 clustering changes the mechanism of the receptor endocytosis by engaging dynamin-2-dependent CIE pathways. b The chemical structure of monomethyl auristatin E bearing the valine-citrulline linker (vcMMAE). c The schematic representation of the conjugation of T-Fc with the cytotoxic compound MMAE. The Fc region of IgG (CH2 and CH3 domains) is labeled in gray, and anti-FGFR1 scFv proteins (VH and VL fusions) are marked in blue. Antibody regions recognizing epitopes within FGFR1 are marked in orange. Thiol groups of reduced cysteines are marked in yellow and attached cytotoxic payloads are marked in red. d , e The efficiency of the conjugation and purity of T-Fc-vcMMAE were analyzed with SDS/PAGE ( d ) and western blotting ( e ) with antibodies recognizing the Fc fragment. f The spectroscopic analysis of DAR parameter for T-Fc-vcMMAE. DAR was calculated through the absorbance measurement for T-Fc and T-Fc-vcMMAE at 248 nm and 280 nm wavelengths according to (Chen ).

Article Snippet: To examine the interaction of antibody alone and its conjugate with murine FGFR1, each compound (40 nM) was injected on the CM4 sensor chip with immobilized murine recombinant FGFR1 (10,135-FR, R&D Systems) at 535 RU for 120 s at 30 μL·min −1 flow rate.

Techniques: Conjugation Assay, Binding Assay, Activation Assay, Clinical Proteomics, Membrane, Labeling, SDS Page, Western Blot

Journal: Molecular Medicine

Article Title: The cytotoxic conjugate of highly internalizing tetravalent antibody for targeting FGFR1-overproducing cancer cells

doi: 10.1186/s10020-021-00306-2

Figure Lengend Snippet: Interaction of T-Fc-vcMMAE with FGFR1. a Evaluation of T-Fc and T-Fc-vcMMAE interaction with FGFR1 by BLI. The extracellular region of FGFR1 was immobilized on BLI sensors and incubated either with T-Fc or T-Fc-vcMMAE. The association and dissociation profiles were measured. b SPR-determined kinetic parameters of the interaction between T-Fc-vcMMAE and FGFR1. The extracellular region of FGFR1 was immobilized on SPR sensors and incubated with various concentrations of T-Fc-vcMMAE. K D value is presented. c SPR results of the interaction between T-Fc and T-Fc-vcMMAE, and murine FGFR1, respectively. The murine recombinant FGFR1 was immobilized on SPR sensors and incubated with T-Fc or T-Fc-vcMMAE. The association and dissociation profiles were measured. d T-Fc and T-Fc-vcMMAE are unable to activate FGFR1. Serum-starved NIH3T3 cells were incubated with FGF1 (positive control) or with different concentrations of T-Fc or T-Fc-vcMMAE. Cells were lysed and activation of FGFR1, and receptor-downstream signaling was assessed with western blotting (WB). The level of tubulin served as a loading control

Article Snippet: To examine the interaction of antibody alone and its conjugate with murine FGFR1, each compound (40 nM) was injected on the CM4 sensor chip with immobilized murine recombinant FGFR1 (10,135-FR, R&D Systems) at 535 RU for 120 s at 30 μL·min −1 flow rate.

Techniques: Incubation, Recombinant, Positive Control, Activation Assay, Western Blot, Control

Journal: Molecular Medicine

Article Title: The cytotoxic conjugate of highly internalizing tetravalent antibody for targeting FGFR1-overproducing cancer cells

doi: 10.1186/s10020-021-00306-2

Figure Lengend Snippet: FGFR1-mediated internalization of T-Fc and T-Fc-vcMMAE. a FGFR1-dependent endocytosis of T-Fc and T-Fc-vcMMAE. U2OS-R1 cells stably expressing FGFR1 were incubated with T-Fc or T-Fc-vcMMAE for 15 min at 37 °C. Nuclei were stained with NucBlue Live, and early endosomes were labeled by CellLight Early Endosomes-RFP. Cells were fixed, and internalized antibodies were visualized with Zenon AF-488 using wide-field fluorescence microscope. Scale bar represents 20 μm. b – e Confocal microscopy analysis of the T-Fc and T-Fc-vcMMAE internalization. B and C . U2OS-R1 cells were briefly incubated with T-Fc and T-Fc-vcMMAE (t = 0 min) and analyzed with quantitative immunofluorescence microscopy using Zenon-AF-488 to label Fc-bearing recombinant proteins. d , e T-Fc and T-Fc-vcMMAE were incubated with U2OS-R1 cells for different time periods (5, 15, 30, 60 min) and internalized antibodies were labeled with Zenon AF-488, and analyzed with confocal microscopy. Scale bar represents 50 μm. Quantification of T-Fc and T-Fc-vcMMAE internalization (expressed as integral fluorescence intensity in arbitrary units, AU) was performed using the HARMONY software. Mean values of three independent experiments of integral intensity of Zenon AF-488 signal ( c ) and integral intensity of Zenon AF-488 vesicles ( e ) ± SEM are shown. T -test was used to assess the statistical significance of measured differences in internalization; * p < 0.05, ** p < 0.01, *** p < 0.0001, n.s.- not significant. f Efficiency and selectivity of T-Fc and T-Fc-vcMMAE internalization studied with flow cytometry. Internalization was analyzed with serum-starved U2OS and U2OS-R1 cells, treated with T-Fc or T-Fc-vcMMAE labeled with DyLight550. After 40 min incubation on ice, cells were transferred to 37 °C for 15 min, and then subsequently analyzed by flow cytometry

Article Snippet: To examine the interaction of antibody alone and its conjugate with murine FGFR1, each compound (40 nM) was injected on the CM4 sensor chip with immobilized murine recombinant FGFR1 (10,135-FR, R&D Systems) at 535 RU for 120 s at 30 μL·min −1 flow rate.

Techniques: Stable Transfection, Expressing, Incubation, Staining, Labeling, Fluorescence, Microscopy, Confocal Microscopy, Immunofluorescence, Recombinant, Software, Flow Cytometry

Journal: Molecular Medicine

Article Title: The cytotoxic conjugate of highly internalizing tetravalent antibody for targeting FGFR1-overproducing cancer cells

doi: 10.1186/s10020-021-00306-2

Figure Lengend Snippet: Cytotoxicity of T-Fc-vcMMAE against FGFR1-overproducing cancer cells. a FGFR1 expression levels in studied cell lines analyzed by western blotting using anti-FGFR1 antibody. Tubulin level assessed with anti-tubulin antibody served as a loading control. b – i Cytotoxic potential of T-Fc and T-Fc-vcMMAE. Negative-FGFR1 cells: U2OS ( b ), HCC15 ( d ) and T47D ( e ), and positive-FGFR1 cells: U2OS-R1 ( c ), JIMT-1 ( f ), COLO-699 ( g ), NCI-H520 ( h ) and NCI-H1581 ( i ) were treated with indicated agents at various concentrations for 96 h and their viability was assessed with the Presto Blue assay. Presented results are mean values from three experiments ± SD. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.0001, not significant differences are not marked. j EC 50 values for T-Fc-vcMMAE for each cell line, respectively. EC50 values were calculated based on the Hill equation using Origin 7 software (Northampton, MA). k The T-Fc internalization into studied cancer cell lines, visualized with fluorescence microscopy. Cell line tested in cytotoxic assays were incubated with 15 µg/mL of T-Fc for 30 min at 37 °C. Nuclei were stained with NucBlue Live, cells were fixed, and internalized antibodies were visualized with Zenon AF-488 using wide-field fluorescence microscopy. Scale bar represents 20 μm

Article Snippet: To examine the interaction of antibody alone and its conjugate with murine FGFR1, each compound (40 nM) was injected on the CM4 sensor chip with immobilized murine recombinant FGFR1 (10,135-FR, R&D Systems) at 535 RU for 120 s at 30 μL·min −1 flow rate.

Techniques: Expressing, Western Blot, Control, Software, Fluorescence, Microscopy, Incubation, Staining

Journal: Nature Communications

Article Title: FGF4-FGFR1 signaling promotes podocyte survival and glomerular function to ameliorate diabetic kidney disease in male mice

doi: 10.1038/s41467-025-65978-4

Figure Lengend Snippet: a Relative mRNA levels of Fgfr1-4 in total renal tissue lysates from C57BL/6 mice ( n = 16). b , c Immunofluorescence images depicting FGFR1 distribution (green) in normal renal tissues from ( b ) C57BL/6 mice and ( c ) human samples. White dashed lines delineate glomerular boundaries. d , e High-magnification images showing co-localization of FGFR1 (green) with the podocyte marker podocin (red) in ( d ) mouse and ( e ) human glomeruli. f Schematic of podocyte-specific Fgfr1 knockout ( Fgfr1 -PKO), and Western blot of isolated glomeruli confirms loss of FGFR1 in Fgfr1 -PKO versus littermate controls. g , h Changes in ( g ) blood glucose levels and ( h ) BUN and UACR across treatment groups ( n = 6). i Renal excretion kinetics, GFR, and elimination half-life ( t 1/2 ) in mice from each group ( n = 3). j , k Histological examination and quantification of renal sections using H&E, PAS, and Masson’s trichrome staining ( n = 6). l , m immunohistochemistry and quantification of renal sections for the podocyte marker Nephrin from the indicated groups ( n = 6). n , o Representative images of isolated mouse glomeruli stained with WT-1 (green), TUNEL (green), and DHE (red), with quantification shown in ( o ) ( n = 6). Nuclei were counterstained with DAPI (blue). Data are presented as mean ± s.e.m. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 as determined by ordinary one-way ANOVA followed by Tukey’s multiple comparisons test ( a − o ); ns, not significant.

Article Snippet: Fgf4- floxed mice ( Fgf4 flox/flox , Cat. No. T009264) were obtained from GemPharmatech Co., Ltd. (Jiangsu, China), while Fgfr1- flox mice ( Fgfr1 flox/flox ) and podocin (Pod) -Cre mice were acquired from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Immunofluorescence, Marker, Knock-Out, Western Blot, Isolation, Staining, Immunohistochemistry, TUNEL Assay

Journal: Nature Communications

Article Title: FGF4-FGFR1 signaling promotes podocyte survival and glomerular function to ameliorate diabetic kidney disease in male mice

doi: 10.1038/s41467-025-65978-4

Figure Lengend Snippet: a , b Human podocytes were isolated from the urine of DKD patients and exposed to high glucose conditions with or without rFGF4 treatment. a Representative images of TUNEL staining (green) and quantification of TUNEL-positive cells from the indicated groups ( n = 6). b Immunofluorescence analysis of tubulin (green) and quantification of the aspect ratio (width to height ratio) of human podocytes ( n = 6). White dashed lines mark the edge of the cells. c Representative images and quantification of isolated human glomeruli stained for WT-1 (green), TUNEL (green), DHE (red), and Nrf-2 (pink). Nuclei were counterstained with DAPI (blue). d Western blot and quantitative analysis of c-Cas3, CAT, SOD-2, and Nrf-2 protein levels in human glomeruli from the indicated groups ( n = 4). β-actin served as a loading control. e Western blot analysis of the activation of AMPK-FOXO1 pathway components in total glomerular tissue lysate. The ratios of phosphorylated proteins to total proteins are shown as scatter plots ( n = 4). f Schematic showing that FGF4 protects against DKD by activating the FGFR1-AMPK-FOXO1 signaling axis. Data are presented as mean ± s.e.m. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 as determined by unpaired two-tailed Student’s t -test ( a , b ) or ordinary one-way ANOVA followed by Tukey’s multiple comparisons test ( c − e ); ns, not significant. c-Cas3, cleaved-Caspase3; CAT, Catalase.

Article Snippet: Fgf4- floxed mice ( Fgf4 flox/flox , Cat. No. T009264) were obtained from GemPharmatech Co., Ltd. (Jiangsu, China), while Fgfr1- flox mice ( Fgfr1 flox/flox ) and podocin (Pod) -Cre mice were acquired from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Isolation, TUNEL Assay, Staining, Immunofluorescence, Western Blot, Control, Activation Assay, Two Tailed Test

Journal: Investigative Ophthalmology & Visual Science

Article Title: Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro

doi: 10.1167/iovs.61.12.27

Figure Lengend Snippet: FGFR, Fibroblast Growth Factor Receptor

Article Snippet: The following Abs were used: mouse anti-cytokeratin-3/2p monoclonal antibody (mAb; AE-5, 1:100; Santa Cruz Biotechnology, Inc, Dallas, TX, USA), mouse anti-human cytokeratin-4 mAb (6B10, 1:300; Abcam, Cambridge, UK), rabbit monoclonal [EP1599Y] anti-human cytokeratin 4 (1:200, Abcam), mouse anti-human cytokeratin-7 mAb (RCK105, 1:10000; Millipore, Billerica, MA, USA), mouse anti-human cytokeratin-13 mAb (1:20; American Research Products, Inc., Palos Verdes, CA, USA), mouse anti-human cytokeratin-12 mAb (N-16; 1:500; Santa Cruz Biotechnology), rabbit anti-cytokeratin 12/K12 mAb (1:200, EPR17882, Abcam), rabbit anti-human MUC5AC polyclonal Ab (H-160, 1:100, Santa Cruz Biotechnology), anti-FGFR1 mAb (1:200, 133105; R&D Systems, Minneapolis, MN, USA), anti-human FGFR2 (blocking) mAb (1:200, 98725; R&D Systems), anti-FGFR3 mAb (1:200, 135334; R&D Systems), anti-FGFR4 mAb (1:200, 137114; R&D Systems), and anti-FGFR1 blocking mAb (VBS1; Chemicon, Temecula, CA, USA).

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro

doi: 10.1167/iovs.61.12.27

Figure Lengend Snippet: FGF receptor expression and effect of FGF receptor blockade in colonies derived from adherent single cells. ( A ) FGFR1 and FGFR2, but not FGFR3 and FGFR4, are detected in the adherent colonies by RT-PCR. ( B ) FGFR1 and FGFR2 expression is detected by Western blotting. ( C ) Effect of FGF receptor blockade was tested in colonies derived from adherent single cells. PAS-positive colony number in the anti-FGFR1 monoclonal antibody group significantly decreases as compared with that in the control Ig group. Treatment with an anti-FGFR2 blocking mAb does not affect the number of colony. ( D ) Colony with amorphous material and epithelium is observed with control IgG. ( E ) Anti-FGFR1 mAb suppresses the proliferation of both goblet-like cell and epithelium. Similar results were obtained with repeated three experiments. M, size markers; S, sample; P, positive control; N, negative control, N.S., not significant.

Article Snippet: The following Abs were used: mouse anti-cytokeratin-3/2p monoclonal antibody (mAb; AE-5, 1:100; Santa Cruz Biotechnology, Inc, Dallas, TX, USA), mouse anti-human cytokeratin-4 mAb (6B10, 1:300; Abcam, Cambridge, UK), rabbit monoclonal [EP1599Y] anti-human cytokeratin 4 (1:200, Abcam), mouse anti-human cytokeratin-7 mAb (RCK105, 1:10000; Millipore, Billerica, MA, USA), mouse anti-human cytokeratin-13 mAb (1:20; American Research Products, Inc., Palos Verdes, CA, USA), mouse anti-human cytokeratin-12 mAb (N-16; 1:500; Santa Cruz Biotechnology), rabbit anti-cytokeratin 12/K12 mAb (1:200, EPR17882, Abcam), rabbit anti-human MUC5AC polyclonal Ab (H-160, 1:100, Santa Cruz Biotechnology), anti-FGFR1 mAb (1:200, 133105; R&D Systems, Minneapolis, MN, USA), anti-human FGFR2 (blocking) mAb (1:200, 98725; R&D Systems), anti-FGFR3 mAb (1:200, 135334; R&D Systems), anti-FGFR4 mAb (1:200, 137114; R&D Systems), and anti-FGFR1 blocking mAb (VBS1; Chemicon, Temecula, CA, USA).

Techniques: Expressing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Blocking Assay, Positive Control, Negative Control