Journal: bioRxiv

Article Title: An FGF7-FGFR2-KLF4 feedback loop sustains anti-inflammatory signaling in epithelial cells

doi: 10.64898/2026.03.20.711763

Figure Lengend Snippet: A) RT-qPCR for IL6 relative to RPL27 using RNA from serum-starved HaCaT keratinocytes, which had been pre-treated for 1 h with actinomycin D or vehicle and incubated for 6 h with FGF7 or vehicle (N = 12 per treatment group). B) RT-qPCR for IL6 using RNA from serum-starved HaCaT keratinocytes, pre-treated for 2 h with LY294002 (PI3K inhibitor), U0126 (MEK1/2 inhibitor) or vehicle, followed by 6 h FGF7 treatment (N = 6). C) RT-qPCR for IL6 using RNA from HaCaT keratinocytes, treated for 6 h with the FGFR kinase inhibitors BGJ398 (N = 9), AZD4547 (N = 3) or erdafitinib (N = 3) in 10% FBS. D) RT-qPCR for IL6 using RNA from serum-starved WT and FGFR2 KO HaCaT cell lines, treated for 6 h with FGF7 or vehicle (N = 9; 3 different WT and KO cell lines). E) RT-qPCR for IL6 using RNA from serum-starved HaCaT or human primary keratinocytes (HPKs), pre-treated for 3 h with FGF7 or vehicle and incubated for 3 h with poly(I:C) or vehicle (N = 6; HPKs from two donors). F) RT-qPCR for IL6 using RNA from serum-starved HaCaT keratinocytes, pre-treated for 3 h with FGF7 or vehicle and incubated for 3, 6 or 12 h with TNFα or vehicle (N = 3). G) Relative IL-6 levels (ELISA) in conditioned medium from serum-starved HaCaT keratinocytes, pre-treated for 3 h with FGF7 or vehicle and incubated for 6 or 12 h with TNFα or vehicle (N = 3). H) IL6 promoter cloning strategy showing 2044 or 1057 base pair (bp) fragments including the transcription start site (TSS), inserted into a firefly luciferase vector. I) Luciferase activity in lysates of HaCaT keratinocytes, stably transduced with lentiviruses containing the long or short IL6 promoter fragment in front of the luciferase gene. Cells were serum-starved and treated for 6 h with FGF7 or vehicle (N = 6). J, K) Luciferase activity in lysates of HaCaT keratinocytes, stably transduced with lentiviruses containing the long or short IL6 promoter fragment in front of the luciferase gene. Cells had been serum-starved and pre-treated for 3 h with FGF7 or vehicle and incubated for 6 h with poly(I:C), TNFα, or vehicle (N = 6). Data information: Graphs show mean and standard deviation (SD). ns: non-significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Mann-Whitney U test (A, B, I, normalized to respective control), one-way ANOVA with Bonferroni’s multiple comparisons test (C), 2-way ANOVA with Bonferroni’s multiple comparisons test (D; E, normalized to respective control; F; J; K), or Student’s t-test (G, normalized to respective control)).

Article Snippet: Cells were treated with human FGF7 or FGF10 (10 ng/ml, both from Peprotech, Rocky Hill, NJ), poly(I:C) (1 μg/ml, InvivoGen, San Diego, CA), TNFα (20 ng/ml, Peprotech), 2’3’-cGAMP (10 μg/ml, InvivoGen), 3p-hpRNA (1 μg/ml, InvivoGen), actinomycin D (1 μg/ml; Sigma-Aldrich, St. Louis, MO), the FGFR kinase inhibitors BGJ398 (3.6 μM), AZD4547 (10 μM) or erdafitinib (5 μM) (all from Selleckchem, Frankfurt, Germany), the MEK1/2 inhibitor U0126 (10 μM; Calbiochem), the PI3K inhibitor LY294002 (5 μM; Calbiochem), FITC-LC-TAT (100 nM, AnaSpec, Fremont, CA) and/or KLF4-TAT (100 nM, Peprotech).

Techniques: Quantitative RT-PCR, Incubation, Enzyme-linked Immunosorbent Assay, Cloning, Luciferase, Plasmid Preparation, Activity Assay, Stable Transfection, Transduction, Standard Deviation, MANN-WHITNEY, Control

Journal: Nutrients

Article Title: Synergistic Anticancer Effects of Fibroblast Growth Factor Receptor Inhibitor and Cannabidiol in Colorectal Cancer

doi: 10.3390/nu17162609

Figure Lengend Snippet: Confirmation of FGFR expression, and evaluation of FGFR inhibitor and CBD effects in CRC lines. ( A ) Endogenous protein levels of FGFR family in various cancer cell lines were determined by Western blot analysis. β-actin was used as a loading control. ( B ) Colorectal cancer and normal colon cell lines were treated with AZD4546 and CBD for 24 h. Cell viability was measured by WST-1 assay. ( C ) NCI-H716 cells were treated with increasing concentrations of CBD and AZD4547 for 24 h, and cell proliferation was measured by WST-1 assay. ( D ) Western blot analysis of NCI-H716 cells treated with AZD4547 (10 nM) for 24 h, showing inhibition of FGFR downstream signaling pathways. β-actin was used as a loading control.

Article Snippet: FGFR inhibitors (AZD4547, PD173074 , BGJ398) were purchased from Selleckchem (Houston, TX, USA).

Techniques: Expressing, Western Blot, Control, WST-1 Assay, Inhibition, Protein-Protein interactions

Journal: Hepatology International

Article Title: Fibroblast growth factor inhibition by molecular-targeted agents mitigates immunosuppressive tissue microenvironment in hepatocellular carcinoma

doi: 10.1007/s12072-023-10603-z

Figure Lengend Snippet: Tumor immune microenvironment is altered by FGFR-1/2/3/4 inhibitor treatment. Representative micrographs (upper panel) and quantification (lower panel) of CD3-positive, CD8-positive, Granzyme B-positive, Foxp3-positive, F4/80-positive areas, Arginase-1-positive areas, CD11c-positive, NK1.1-positive, PD-1-, and PD-L1-positive cells in the VT and FGFR inhibitor groups, respectively. Arrowheads represent positive cells. * p < 0.05, ** p < 0.01 vs. VT, Student t test. Data are presented as mean ± SEM. VT vehicle treatment, FGFR fibroblast growth factor receptor, ns not significant, PD-1 programmed cell death-1, PD-L1 programmed cell death ligand 1

Article Snippet: The treatment groups were as follows: (I) Sorafenib; sorafenib tosylate (CAS No.: 475207-59-1; purchased from Santa Cruz Biotechnology, Inc., TX, USA), administered 30 mg/kg orally/once daily; (II) Lenvatinib; lenvatinib mesylate (CAS No.: 417716-92-8; kindly provided by Eisai co. ltd., Tokyo, Japan), administered 10 mg/kg/mouse orally/once daily; (III) VEGFR2 (VEGF receptor 2) antibody; DC101, an anti-VEGFR2 antibody, purchased from Bio X Cell (Lebanon, NH, USA), administered intraperitoneally thrice/week at a dose of 40 mg/kg; (IV) Regorafenib; regorafenib (CAS No.: 755037-03-7; purchased from Tokyo Chemical Industry, Tokyo, Japan), administered 3 mg/kg orally/once daily; (V) Cabozantinib; cabozantinib malate (CAS No.: 849217-68-1; purchased from ChemScene, Monmouth Junction, NJ, USA), administered 30 mg/kg orally/once daily; (VI) FGFR (FGF receptor) inhibitor; AZD4547, FGFR-1/2/3/4 inhibitor (CAS No.: 1035270-39-3; purchased from ChemScene), administered 12.5 mg/kg intraperitoneally/once daily; (VII) VT (vehicle treatment); PBS as vehicle was administered orally or intraperitoneally.

Techniques: