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