recombinant human fgf1  (R&D Systems)

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A) ETV4 expression in breast tumors and paired normal breast tissue from people with breast cancer. Data obtained from tnmplot.com. Mann-Whitney test determined significance. (B) High ETV4 expression (RNA seq) associates with lower overall survival for people with breast cancer. HR=1.44 (1.14-1.82). (C) Tumors with high ETV4 expression (mRNA z-score >1.5) are more likely to be ER-negative than ER-positive. Chi-squared test q-value <0.0001. (D) High ETV4 expression (RNA seq) associates with lower overall survival for patients with ER-positive breast cancer. HR=1.39 (1.05-1.85). (E) High ETV4 expression (array) associates with lower recurrence-free survival for patients with lymph node-positive ER-positive breast cancer. HR=1.3 (1.031-1.628). (F) ETV4 expression in ER-positive breast tumors stratified by pathologic complete response after aromatase inhibitors using data from ROCplotter.com. (G) ETV4 expression in ER-positive tumors stratified by pathologic complete response to any chemotherapy using data from ROCplotter.com. Mann-Whitney test determined significance for f and g . (H) ETV4 expression (RNA seq) in UCD12 PDX tumors from lean or obese female mice. (I) ETV4 expression (array) in tumors from patients with ER-positive breast cancer (data from GSE24185). (J) Pearson correlation between ETV4 and FGF1 expression in tumors from patients classified as obese, overweight, or lean based on BMI (data from GSE24185). (K) Expression of ETV4 in ER-positive breast cancer cells with or without FGF1 treatment. MCF7 Parental (M7P) or TAMR (M7T); UCD12 (U12). (L) Representative western blot showing ETV4 expression in ER-positive breast cancer cells with or without FGF1 treatment.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Expressing, MANN-WHITNEY, RNA Sequencing, Western Blot

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A) Q-PCR ( left ) and immunoblot ( right ) analysis of ETV4 in MCF7-P cells. (B) Q-PCR ( left ) and immunoblot ( right ) analysis of ETV4 in MCF7-TAMR cells. (C) Representative images of the final timepoint (left) and growth rates (right) of MCF7-P control or ETV4-knockdown cells treated with vehicle or FGF1. (D) Area confluence relative to control vehicle time 0 of cells at the final timepoint following treatment of MCF7-P cells. Two-way ANOVA testing for main effects of ETV4 knockdown or FGF1 treatment or interaction was performed. P-values indicate post-hoc multiple testing for specific differences between pre-defined comparisons. (E) Representative images of the final timepoint (left) and growth rates (right) of MCF7-TAMR control or ETV4-knockdown cells treated with vehicle or FGF1. (F) Area confluence relative to control vehicle time 0 of cells at the final timepoint following treatment of MCF7-TAMR cells. Two-way ANOVA testing for main effects of ETV4 knockdown or FGF1 treatment or interaction was performed. P-values indicate post-hoc multiple testing for specific differences between pre-defined comparisons. (G) Dose-response curve and interpolated IC 50 values of tamoxifen treatment in MCF7-P control and ETV4 knockdown cells following treatment with or without FGF1. (H) Dose-response curve and interpolated IC 50 values of tamoxifen treatment in MCF7-TAMR control and ETV4 knockdown cells following treatment with or without FGF1.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Western Blot, Control, Knockdown

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A) Expression of ETV4 gene ( left ) and protein ( right ) following knockdown in MCF7-P cells. (B) Expression of ETV4 gene ( left ) and protein ( right ) following knockdown in MCF7-TAMR cells. (C) Representative images of the final timepoint (left) and growth rates (right) of MCF7-P control or ETV4-overexpressing cells treated with vehicle or FGF1. (D) Area confluence relative to control vehicle time 0 of cells at the final timepoint following treatment of MCF7-P cells. Two-way ANOVA testing for main effects of ETV4 overexpression or FGF1 treatment or interaction was performed. P-values indicate post-hoc multiple testing for specific differences between pre-defined comparisons. (E) Representative images of the final timepoint (left) and growth rates (right) of MCF7-TAMR control or ETV4-overexpressing cells treated with vehicle or FGF1. (F) Area confluence relative to control vehicle time 0 of cells at the final timepoint following treatment of MCF7-TAMR cells. Two-way ANOVA testing for main effects of ETV4 overexpression or FGF1 treatment or interaction was performed. P-values indicate post-hoc multiple testing for specific differences between pre-defined comparisons. (G) Dose-response curve and interpolated IC 50 values of tamoxifen treatment in MCF7-P control and ETV4 overexpressing cells following treatment with or without FGF1. (H) Dose-response curve and interpolated IC 50 values of tamoxifen treatment in MCF7-TAMR control and ETV4 overexpressing cells following treatment with or without FGF1. (I) Dose-response curve and interpolated IC 50 values of BGJ398 treatment in MCF7-P and MCF7-TAMR control and ETV4 overexpressing cells.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Expressing, Knockdown, Control, Over Expression

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A) Bubble plot of gene set enrichment analysis (GSEA) showing enriched pathways in MCF7-TAMR control, ETV4 knockdown, and ETV4-overexpressing cells with or without FGF1 stimulation. (B) Hallmarks of Cancer enrichment plot illustrating the pathways represented by genes that are downregulated by ≥2-fold (adjusted p-value) in ETV4 knockdown compared with control vehicle-treated MCF7-TAMR cells. Bar height reflects −log10 adjusted p-value, with dashed circles indicating significance thresholds. (C) Volcano plot comparing ETV4 knockdown versus control vehicle-treated MCF7-TAMR cells, highlighting downregulated genes involved in reprogramming energy metabolism. Blue points denote significantly downregulated genes. Blue points denote significantly downregulated genes that correspond to the significant cancer hallmarks. (D) Hallmarks of Cancer enrichment plot illustrating the pathways represented by genes that are upregulated by ≥1.58-fold (adjusted p-value) in MCF7-TAMR ETV4 overexpressing cells treated with FGF1 vs vehicle controls. Bar height reflects −log10 adjusted p-value, with dashed circles indicating significance thresholds, including sustaining proliferative signaling. (E) Volcano plot comparing genes significantly altered in MCF7-TAMR ETV4 overexpressing cells treated with or without FGF1. Red points denote significantly upregulated genes that correspond to the significant cancer hallmarks.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Control, Knockdown

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A-B) Heatmaps showing mRNA expression levels of glycolytic pathway genes (HK2, PFKP, PGK1, ENO1, and LDHA) under vehicle and FGF1-treated conditions in MCF7-P ( a ) and MCF7-TAMR ( b ) control and ETV4 knockdown cells, respectively. Data are expressed as fold change versus the average of vehicle treated cells for each gene, showing 3 replicates per group. (C) Seahorse metabolic flux analysis showing the kinetic graph of oxygen consumption rate (OCR) in MCF7-P cells. (D-G) Metabolic parameters including basal respiration ( d ), maximal respiration ( e ), ATP-production coupled respiration ( f ), and ECAR ( g ) in control and ETV4 knockdown MCF7-P cells, upon vehicle and FGF1 stimulation. Data analyzed using a 2-way ANOVA testing for main effects of ETV4 or FGF1 treatment or interactions. P-values denote post-hoc analysis of specific comparisons. (H) Seahorse metabolic flux analysis showing the kinetic graph of oxygen consumption rate (OCR) in MCF7-TAMR cells. (I-L) Metabolic parameters including basal respiration ( i ), maximal respiration ( j ), ATP-production coupled respiration ( k ), and ECAR ( l ) in control and ETV4 knockdown MCF7-TAMR cells, upon vehicle and FGF1 stimulation. Data were analyzed using a 2-way ANOVA testing for main effects of ETV4 or FGF1 treatment or interactions. P-values denote post-hoc analysis of specific comparisons. All Seahorse data were normalized to total protein in each well. N=16-24 replicates per measure.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Expressing, Control, Knockdown

Journal: bioRxiv

Article Title: The Ets transcription factor ETV4 regulates FGF1-dependent proliferation and glycolysis in ER-positive breast cancer

doi: 10.64898/2026.01.13.699240

Figure Lengend Snippet: (A-B) Heatmaps showing mRNA expression levels of glycolytic pathway genes (HK2, PFKP, PGK1, ENO1, and LDHA) under vehicle and FGF1-treated conditions in MCF7-P ( a ) and MCF7-TAMR ( b ) control and ETV4 overexpressing cells, respectively. Data are expressed as fold change versus the average of vehicle treated cells for each gene, showing 3 replicates per group. (C) Seahorse metabolic flux analysis showing the kinetic graph of oxygen consumption rate (OCR) in MCF7-P cells. (D-G) Metabolic parameters including basal respiration ( d ), maximal respiration ( e ), ATP-production coupled respiration ( f ), and ECAR ( g ) in control and ETV4 overexpressing MCF7-P cells upon vehicle and FGF1 stimulation. Data were analyzed using a 2-way ANOVA testing for main effects of ETV4 or FGF1 treatment or interactions. P-values denote post-hoc analysis of specific comparisons. (H) Seahorse metabolic flux analysis showing the kinetic graph of oxygen consumption rate (OCR) in MCF7-TAMR cells. (I-L) Metabolic parameters including basal respiration ( i ), maximal respiration ( j ), ATP-production coupled respiration ( k ), and ECAR ( l ) in control and ETV4 knockdown MCF7-TAMR cells, upon vehicle and FGF1 stimulation. Data were analyzed using a 2-way ANOVA testing for main effects of ETV4 or FGF1 treatment or interactions. P-values denote post-hoc analysis of specific comparisons. All Seahorse data were normalized to total protein in each well. N=16-24 replicates per measure.

Article Snippet: Recombinant human FGF1 was purchased from R&D Systems and was diluted in 0.01% bovine serum albumin (BSA) in PBS and used at a final concentration of 5 ng/mL.

Techniques: Expressing, Control, Knockdown

Journal: Molecular Metabolism

Article Title: Sustained diabetes remission induced by FGF1 involves a shift in transcriptionally distinct AgRP neuron subpopulations

doi: 10.1016/j.molmet.2025.102300

Figure Lengend Snippet: snRNA-seq study design and identified cell populations . A , FGF1 or saline was injected icv to Lep ob/ob or C57BL/6 mice. BL6 and the group of Lep ob/ob mice receiving saline were pair-fed to the FGF1 injected Lep ob/ob mice (FGF1) for up to seven days after injection. Mediobasal hypothalamus samples for snRNA-seq were collected five or 14 days after injections and nuclei were sorted for snRNA-seq. B,C, UMAP plot of neuronal (left) and non-neuronal (right) cells across day five and 14 following label transfer from using data from Campbell et al. and Affinati et al. . Abbreviations: FGF1, fibroblast growth factor 1; icv, intracerebroventricular injection; UMAP, uniform manifold approximation and projection.

Article Snippet: After one week of recovery, mice were injected with 1.5 μL of either recombinant human FGF1 (2 μg/μL, Novo Nordisk) or saline solution using a syringe pump at 2 μL/min.

Techniques: Saline, Injection

Journal: Molecular Metabolism

Article Title: Sustained diabetes remission induced by FGF1 involves a shift in transcriptionally distinct AgRP neuron subpopulations

doi: 10.1016/j.molmet.2025.102300

Figure Lengend Snippet: FGF1 induces cell type-specific shifts in hypothalamic transcriptional neighborhoods at Day 5 . A, UMAP of all neurons highlighting cells from FGF1-ob (black) and Veh-ob (gray) mice. UMAP of all neuronal cells (upper panel) and non-neurons (lower panel) B, UMAP highlighting cells mapped to FGF1-enriched (red) and FGF1-depleted (blue) neighborhoods. C , Beeswarm plots of all neighborhoods across all cell populations with ≥20% regulated neighborhoods D , Percent FGF1-enriched (red) or FGF1-depleted (blue) neighborhoods out of the total number of neighborhoods in a given cell population Abbreviations: FGF1, fibroblast growth factor 1; icv, intracerebroventricular injection; UMAP, uniform manifold approximation and projection; FGF1-ob, Lep ob/ob mice treated with FGF1 via icv injection; Veh-ob, Lep ob/ob mice injected with saline and pair-fed to match food intake of FGF1-ob animals; Veh-WT, wild-type C57BL/6 mice injected with saline and pair-fed to match food intake of FGF1-ob animals; FGF1-depleted, neighborhoods in which the abundance of FGF1-ob cells is decreased relative to Veh-ob cells; FGF1-enriched, neighborhoods in which the abundance of FGF1-ob cells is increased relative to Veh-ob cells. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After one week of recovery, mice were injected with 1.5 μL of either recombinant human FGF1 (2 μg/μL, Novo Nordisk) or saline solution using a syringe pump at 2 μL/min.

Techniques: Injection, Saline

Journal: Molecular Metabolism

Article Title: Sustained diabetes remission induced by FGF1 involves a shift in transcriptionally distinct AgRP neuron subpopulations

doi: 10.1016/j.molmet.2025.102300

Figure Lengend Snippet: Cell populations exhibiting transcriptional changes towards wild-type phenotypes five days after icv FGF1 injection . A, UMAP of all neurons highlighting vehicle-treated cells across Lep ob/ob and wild-type strains. B , UMAP of all neurons highlighting cells mapped to WT-enriched (magenta) and WT-depleted (green) neighborhoods. C , Percent WT-enriched or WT-depleted neighborhoods out of the total number of neighborhoods incell populations with at least 25% of NH abundance changed. D , Schematic of neighborhood abundance changes in FGF1 KO vs Veh-ob (top) and Veh-WT vs Veh-ob (bottom), illustrating FGF1- and WT-enriched/depleted neighborhoods. Central overlaps represent cells that change in the same direction in both analyses. E, Percent rescued-enriched or rescued-depleted neighborhoods out of the total number of neighborhoods in each cell population. F , Focused view of AgRP Milo neighborhoods in UMAP space. Each neighborhood is color-coded based on whether its cell abundance was significantly altered by icv FGF1 treatment, by genotype (Lep ob/ob vs. WT), and whether these changes reflect a rescue toward the wild-type state following FGF1 administration. G , Gene Ontology enrichment results depicting the most enriched terms for genes in depleted- and enriched rescued AgRP neighborhoods. Abbreviations: FGF1, fibroblast growth factor 1; icv, intracerebroventricular injection; UMAP, uniform manifold approximation and projection; FGF1-ob, hyperglycemic Lep ob/ob treated with FGF1 via icv injection; Veh-ob, Lep ob/ob mice injected with saline and pair-fed to match food intake of FGF1-ob animals; WT, wild-type animals injected with saline and pair-fed to match food intake of FGF1-ob animals; FGF1-enriched, neighborhoods in which the abundance of FGF1-ob cells is increased relative to Veh-ob cells; FGF1-depleted, neighborhoods in which the abundance of FGF1-ob cells is decreased relative to Veh-ob cells; WT-enriched, neighborhoods in which the abundance of WT cells is increased relative to Veh-ob cells; WT-depleted, neighborhoods in which the abundance of WT cells is decreased relative to Veh-ob cells; rescued-enriched, neighborhoods which register a significant differential abundance of FGF1-enriched and WT-enriched; rescued-depleted, neighborhoods that register a significant differential abundance of FGF1-depleted and WT-depleted. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After one week of recovery, mice were injected with 1.5 μL of either recombinant human FGF1 (2 μg/μL, Novo Nordisk) or saline solution using a syringe pump at 2 μL/min.

Techniques: Injection, Saline

Journal: Molecular Metabolism

Article Title: Sustained diabetes remission induced by FGF1 involves a shift in transcriptionally distinct AgRP neuron subpopulations

doi: 10.1016/j.molmet.2025.102300

Figure Lengend Snippet: Gene modules correlated with icv FGF1 injections at Days 5 and 14. A , UMAP plot of AgRP neurons across Days 5 and 14 past icv FGF1 injection, across strains (Lep ob/ob , wild-type) and groups (FGF1-ob, Veh-ob, Veh-WT). Cells from Veh-WT animals differ strongly from cells from leptin-deficient animals (FGF1-ob, Veh-ob). B , Heatmap showing the results of differential module eigengene analysis for FGF1-ob vs. Veh-ob (left) and Veh-WT vs. Veh-ob (right). Colors are scaled by average log 2 fold-change. Significantly different modules were annotated (∗, Bonferroni-adj. p < 0.05). C , UMAP and boxplot of module eigengene scores for the focal WGCNA modules; left UMAP panels show cells colored by their harmonized module eigengene score; boxplots compare the same eigengene scores across WT cells and all FGF1-polarized cells. Polarities significantly different from both FGF1-depleted and ‘Unchanged’ groups were annotated (∗, BH-adj. p < 0.05). D, Top 3 GO term enrichment heatmap for the focal WGCNA modules. E, One-sided Fisher's exact test for enrichment between module hub genes and neuronal activity IEGs. F, Combined module gene network plot (force-directed graph) for the focal WGCNA modules. Abbreviations: FGF1, fibroblast growth factor-1; icv., intracerebroventricular injection; UMAP, Uniform manifold approximation and projection for dimension reduction; FGF1-ob, hyperglycemic Lep ob/ob treated with FGF1 via icv injection; Veh-ob, Lep ob/ob mice injected with saline and pair-fed to match food intake of FGF1-ob animals; WT, wild-type animals injected with saline and pair-fed to match food intake of FGF1-ob animals; GO, gene ontology; BH, Benjamini-Hochberg. IEGs, immediate early genes. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After one week of recovery, mice were injected with 1.5 μL of either recombinant human FGF1 (2 μg/μL, Novo Nordisk) or saline solution using a syringe pump at 2 μL/min.

Techniques: Injection, Activity Assay, Saline

Journal: Molecular Metabolism

Article Title: Sustained diabetes remission induced by FGF1 involves a shift in transcriptionally distinct AgRP neuron subpopulations

doi: 10.1016/j.molmet.2025.102300

Figure Lengend Snippet: Spatial mapping of transcriptomic changes following icv FGF1 injections . A , Brains from four FGF1-ob and four Veh-ob mice were collected five days after injections and cryosectioned for Molecular Cartography. Cell segmentation was used to infer cell boundaries and map transcripts to cells. B , Spatial plot showing the bottom of the third ventricle in the hypothalamus. AgRP neurons with FGF1-depleted or FGF1-enriched polarity are color-coded accordingly. AgRP neurons with the ‘Unchanged’ annotation are coloured black, all other cells are coloured gray. C , Bar plot showing the mean vertical location of FGF1-depleted and FGF1-enriched AgRP neurons relative to ‘Unchanged’ across seven spatial transcriptomics sections. D, Heatmap showing the module assignment and module membership values (kME, eigengene-based connectivity) for perineuronal net genes. E , Chord diagram visualizing the cell–cell communication network of AgRP neurons and non-neuronal cells. F, Heatmap showing the module assignment and module membership values (kME, eigengene-based connectivity) for CellChat genes shown in E. G , Top 3 GO term enrichment heatmap for all available day 5 FGF1-polarities in astrocytes, OPC and oligodendrocytes. H , Following icv injection of FGF1, a subset of transcriptionally distinct, hyperactive AgRP neurons in Lep ob/ob mice transcriptionally shifts toward a dorsal, wild-type-like state by Day 5. This transition is predicted to involve increased GABA A receptor expression, axonogenesis, and enhanced glia–neuron interactions, including perineuronal net remodeling. By Day 14, AgRP neurons exhibit sustained suppression of activity-linked transcriptional programs, consistent with durable neuronal silencing and glycemic remission. Abbreviations: FGF1, fibroblast growth factor 1; UMAP, uniform manifold approximation and projection; icv, intracerebroventricular injection; FGF1-ob, hyperglycemic Lep ob/ob treated with FGF1 via icv injection; Veh-ob, Lep ob/ob mice injected with saline and pair-fed to match food intake of FGF1-ob animals; FGF1-depleted, neighborhoods in which the abundance of FGF1-ob cells is decreased relative to Veh-ob cells; FGF1-enriched, neghborhoods in which the abundance of FGF1-ob cells is increased relative to Veh-ob cells. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After one week of recovery, mice were injected with 1.5 μL of either recombinant human FGF1 (2 μg/μL, Novo Nordisk) or saline solution using a syringe pump at 2 μL/min.

Techniques: Injection, Expressing, Activity Assay, Saline

Journal: Cell Death Discovery

Article Title: FGF1-FGFR2 axis regulated by nuclear receptor RORγ represents an effective strategy in intrahepatic cholangiocarcinoma

doi: 10.1038/s41420-025-02844-8

Figure Lengend Snippet: A Heatmap of mRNA expression changes of FGF family genes and FGFR family genes in HUCCT-1 cells treated for 2 days with 5 μM XY101. B Oncoprint display from cBioPortal of FGF family genes and FGFR family genes alterations in iCCA tumors. C , D qRT-PCR analysis of FGFR2 mRNA in iCCA cells treated with DMSO or RORγ antagonists (GSK805 and XY101) at indicated concentrations for 2 days. n = 3 biological replicates. E qRT-PCR assay of FGF1 mRNA in RBE and HUCCT-1 cells after treatment with DMSO, 5 μM GSK805 or XY101 for 2 days. n = 3 biological replicates. F Immunoblotting assay of indicated proteins in RBE and HUCCT-1 cells after treatment with DMSO, or RORγ antagonists (GSK805 and XY101) at indicated concentrations for 2 days. n = 3 biological replicates. G The genome browser illustrates RORγ-binding events on the promoters of the FGF1 and FGFR2 genes in triple-negative breast cancer cells, as previously reported. These findings are derived from our previous ChIP-seq dataset (GEO: GSE126380 ). H ChIP-qPCR analysis was performed to assess the relative enrichment of RORγ or H3K27ac at the promoters of the FGF1 and FGFR2 genes in iCCA cells treated with 5 μM GSK805 or XY101 for 2 days. The fold change indicates the enrichment of these factors at the gene promoters in response to GSK805 and XY101, normalized to the IgG enrichment in vehicle-treated cells, which was set as 1. All data from in vitro experiments shown above are the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The sources of chemicals are as follows: GSK805, and XY101 were obtained from WuXi AppTec (China); Pemigatinib was purchased from TargetMol (USA); Recombinant human FGF1 was obtained from MedChemExpress (USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Derivative Assay, ChIP-sequencing, ChIP-qPCR, In Vitro

Journal: Cell Death Discovery

Article Title: FGF1-FGFR2 axis regulated by nuclear receptor RORγ represents an effective strategy in intrahepatic cholangiocarcinoma

doi: 10.1038/s41420-025-02844-8

Figure Lengend Snippet: A Immunoblotting analysis of FGF1 protein in iCCA cells transfected with FGF1 siRNAs or control. n = 3 biological replicates. B RBE and HUCCT-1 cells were transfected with FGF1 siRNAs or control. Live cells were counted at the indicated time points after transfection. n = 3 biological replicates. C Transfection of FGF1 siRNAs or control into wild-type or RORγ-overexpressing RBE and HUCCT-1 cells, with or without 5 ng/ml human recombinant FGF1. Viable cells were counted after 4 days. n = 3 biological replicates. D RBE and HUCCT-1 cells were incubated with recombinant human FGF1 at the indicated concentrations for 4 days, after which live cells were counted. n = 3 biological replicates. E Colony formation assays were conducted to detect the survival of RBE and HUCCT-1 cells treated with recombinant human FGF1 at the indicated concentrations for 14 days. n = 3 biological replicates. F FGFR2, phosphorylated FGFR2 (p-FGFR2), and its downstream proteins were detected by immunoblotting in RBE and HUCCT-1 cells treated with recombinant human FGF1 at the specified concentrations for 20 min. n = 3 biological replicates. G Recombinant human FGF1 was added to iCAA cells transfected with RORC siRNAs or control, and live cells were counted after a 4-day incubation. n = 3 biological replicates. H Live cells were counted to detect the survival of RBE and HUCCT-1 cells treated with GSK805, either alone or in combination with human recombinant FGF1 at a concentration of 5 ng/ml. n = 3 biological replicates. I ELISA assay was performed to detect FGF1 levels in the supernatant of RORγ-overexpression and RORC -silenced iCAA cells. n = 3 biological replicates. All data presented above are shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The sources of chemicals are as follows: GSK805, and XY101 were obtained from WuXi AppTec (China); Pemigatinib was purchased from TargetMol (USA); Recombinant human FGF1 was obtained from MedChemExpress (USA).

Techniques: Western Blot, Transfection, Control, Recombinant, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Over Expression