Journal: Biology of reproduction

Article Title: Immunolocalization of acidic and basic fibroblast growth factors in porcine uterine and conceptus tissues.

doi: 10.1095/biolreprod56.6.1527

Figure Lengend Snippet: FIG. 1. Western blot analysis of porcine uterine protein extract at Day 12 of gestation. a) Molecular weight markers (x 10 3) (lane 1), total CM-50 sephadex-extracted uterine protein (25 g, lanes 2, 4, and 6), human recombinant bFGF (100 ng, lanes 3 and 5). Lanes 1 and 2 were stained for total protein by colloidal gold stain. Lanes 3 and 4 were in- cubated with anti-bFGF antibody (0.15 Ixg/ml). Lanes 5 and 6 were in- cubated with rabbit IgG (0.15 .Lg/ml). b) Sephadex-extracted uterine pro- tein (25 jIg, stained for total protein by colloidal gold) in nonreducing 7.5-15% PAGE. Bovine aFGF (500 ng, lanes 2, 4, and 6) and sephadex extract (25 g, lanes 3, 5, and 7). Immunoreactive aFGF bands are ob- served in lanes 2 and 3 (incubated with anti-aFGF antibody). The absence of these bands is noted in lanes 4 and 5 (anti-aFGF antibody immunoab- sorbed with bovine aFGF) and in lanes 6 and 7 (incubated with rabbit IgG).

Article Snippet: For aFGE PAGE was performed using 7.5-15% acrylamide gel and bovine aFGF (cat. #132-FA; R&D Systems, Minneapolis, MN) as standard.

Techniques: Western Blot, Molecular Weight, Recombinant, Staining, Incubation

Journal: British Journal of Cancer

Article Title: Fibroblast growth factor signalling influences homologous recombination-mediated DNA damage repair to promote drug resistance in ovarian cancer

doi: 10.1038/s41416-022-01899-z

Figure Lengend Snippet: A2780DPP cells were lentivirally transduced with an shRNA construct targeting FGF1 or an empty vector control and FGF1 expression assessed using a qRT-PCR analysis and b immunoblot analysis (EV empty vector). MTT assays were used to assess the effect of FGF1 knockdown on sensitivity to c cisplatin (0–25.33 µM) and d carboplatin (0–85.12 µM). FGF1 was overexpressed in A2780 cells using an FGF1-EGFP fusion plasmid and e immunoblot analysis and f green fluorescence imaging used to confirm FGF1 expression. MTT assays were used to assess the influence of FGF1 over-expression on sensitivity to g cisplatin (0–25.33 µM) and h carboplatin (0–85.12 µM). Results represent three independent experiments. Pairwise comparisons of mean IC 50 values and relative gene expression were performed using Student’s t -tests. Scale bar = 100 µm. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: FGF1 (Hs01092738_m1) , ATM (Hs00175892_m1) and 18S ribosomal RNA expression (438839) was assessed in individual 20 μL reactions, combining 10 μL TaqMan universal master mix (Thermo Fisher, Renfrewshire, UK), 1 μL gene-specific probe, 1 μL cDNA and 8 μL nuclease-free water.

Techniques: Transduction, shRNA, Construct, Plasmid Preparation, Control, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Fluorescence, Imaging, Over Expression, Gene Expression

Journal: British Journal of Cancer

Article Title: Fibroblast growth factor signalling influences homologous recombination-mediated DNA damage repair to promote drug resistance in ovarian cancer

doi: 10.1038/s41416-022-01899-z

Figure Lengend Snippet: a The Dundee Ovarian Cancer Study; ‘A1’ sample = chemonaïve, ‘A2’ sample = disease relapse. b FGF1 and FGFR2 gene expression in chemoresistant ovarian cancer patient-derived cells was assessed by qRT-PCR analysis. MTT assays were used to compare viability of chemonaive cells, and cells from drug-resistant relapse samples from two patients in the presence or absence of combination treatments with 10 µM AZD4547 and c , e cisplatin (0–25.33 µM) or d , f carboplatin (0–85.12 µM). Pairwise comparisons of mean IC 50 values and relative gene expression were calculated by Student’s t -tests. * p < 0.05, ** p < 0.01, *** p < 0.001. Ns not significant.

Article Snippet: FGF1 (Hs01092738_m1) , ATM (Hs00175892_m1) and 18S ribosomal RNA expression (438839) was assessed in individual 20 μL reactions, combining 10 μL TaqMan universal master mix (Thermo Fisher, Renfrewshire, UK), 1 μL gene-specific probe, 1 μL cDNA and 8 μL nuclease-free water.

Techniques: Gene Expression, Derivative Assay, Quantitative RT-PCR

Journal: British Journal of Cancer

Article Title: Fibroblast growth factor signalling influences homologous recombination-mediated DNA damage repair to promote drug resistance in ovarian cancer

doi: 10.1038/s41416-022-01899-z

Figure Lengend Snippet: a MTT assays were used to compare viability of A2780, A2780DPP and A2780DPP cells treated with 10 µM KU55933 (ATMi) or 2 µM NU7026 (DNA-PKi) in response to cisplatin (0–25.33 µM). b qRT-PCR analysis was used to assess ATM expression relative to 18S rRNA in response to 5 nM, 10 nM and 20 nM ATM siRNA (±compound SD). c MTT assays were used to compare the viability of A2780, A2780DPP and A2780DPP cells transfected with 20 nM siRNA scrambled negative control or 20 nM si ATM in response to cisplatin (0–25.33 µM). Representative immunoblot analysis comparing the phosphorylation of ATM (S1981) over indicated timepoints following 3 h treatment with 3 µM cisplatin in thymidine synchronised cells (2 mM, 24 h) in d A2780, e A2780DPP and f A2780DPP FGF1 KD cells. A timecourse of FGF1-dependency H2AX (S139; γH2AX) phosphorylation was investigated before ( g – i ) and following treatment with 10 µM KU55933 (ATMi) ( j – l ) in A2780, A2780DPP and A2780DPP FGF1 KD cells. Results are illustrative of four repeat experiments. Pairwise comparisons of mean IC 50 values and relative gene expression were calculated by Student’s t -tests. Scale bar = 100 µm. * p < 0.05, ** p < 0.01, *** p < 0.001. Ns not significant.

Article Snippet: FGF1 (Hs01092738_m1) , ATM (Hs00175892_m1) and 18S ribosomal RNA expression (438839) was assessed in individual 20 μL reactions, combining 10 μL TaqMan universal master mix (Thermo Fisher, Renfrewshire, UK), 1 μL gene-specific probe, 1 μL cDNA and 8 μL nuclease-free water.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Negative Control, Western Blot, Phospho-proteomics, Gene Expression

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Impact of Bmal1 Rescue and Time-Restricted Feeding on Liver and Muscle Proteomes During the Active Phase in Mice

doi: 10.1016/j.mcpro.2023.100655

Figure Lengend Snippet: Bmal1 regulates FGF1, which supports mitochondrial respiration in hepatocytes. A , mRNA ( right ) and protein abundance ( left ) of FGF1 protein in liver. Two-way ANOVA with Tukey’s multiple comparisons test, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001, n = 4. B , Western blot for FGF1 in liver at the indicated diurnal time points. Biological replicates are shown ( bottom ) and quantified ( top ). One-way ANOVA with Fisher’s LSD test, ∗ p < 0.05 and ∗∗ p < 0.01, n = 3. Liver-RE = Bmal1 stopFL/stopFL ; Alfp -Cre tg/0 , hepatocyte-specific reconstitution of Bmal1 . C and D , knockdown or overexpression of Fgf1 in AML12 hepatocytes. si-Ctrl = scrambled sequence; both si-Fgf1 #1 and #2 target Fgf1 but by different sequences. pCMV6-Empty = control vector without Fgf1 open reading frame clone. Left , qPCR, Student’s t test, ∗∗ p < 0.01 and ∗∗∗ p < 0.001, n = 3. Middle , oxygen consumption rate in AML12 hepatocytes. Arrow indicates application of 0.5 μM carbonyl cyanide- p -trifluoromethoxyphenylhydrazone (FCCP) and 1 μM oligomycin. Right , quantification of middle . Basal and stressed values are averages of data points before and after drug application, respectively. Two-way ANOVA with Dunnett’s post hoc test, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001, n = 5 to 6. E , mRNA levels of FGF receptor (FGFR) isoforms in WT (Alfp-Cre+ and Hsa-Cre+) liver. F , measurement of oxygen consumption rate as in D and E . AML12 cells were treated for 24 h with vehicle control (Veh) or the indicated concentration of the pan-FGFR inhibitor AZD4547. Two-way ANOVA with Tukey’s post hoc test, ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001, n = 5 to 6. All mRNA plots are normalized to WT ZT 0, n = 3 per time point per genotype. AL, ad libitum feeding; AML12, alpha mouse liver 12; FGF, fibroblast growth factor; LSD, least significant difference; ns, not significant; qPCR, quantitative PCR; TRF, time-restricted feeding; ZT, zeitgeber time.

Article Snippet: For overexpression experiments, cells were transfected with 100 ng Fgf1 (NM_010197) mouse-tagged ORF Clone (ORIGENE; catalog no.: MR201152) or control empty vector using Lipofectamine 3000 Transfection Reagent (Invitrogen; catalog no.: L3000001) according to the manufacturer’s instructions.

Techniques: Western Blot, Knockdown, Over Expression, Sequencing, Control, Plasmid Preparation, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Cancer

Article Title: Identification of Cathepsin K in the Peritoneal Metastasis of Ovarian Carcinoma Using In-silico, Gene Expression Analysis

doi: 10.7150/jca.14277

Figure Lengend Snippet: Validation of DEGs in POCs versus PMOCs by microfluidic cards Q-PCR.

Article Snippet: Serum concentrations of CTSK, MMP13, and FGF1 were measured by commercial, double-antibody, sandwich ELISA kits (Cusabio Life Science, Wuhan, China).

Techniques: Biomarker Discovery

Journal: Journal of Cancer

Article Title: Identification of Cathepsin K in the Peritoneal Metastasis of Ovarian Carcinoma Using In-silico, Gene Expression Analysis

doi: 10.7150/jca.14277

Figure Lengend Snippet: Immunohistochemistry of representative genes in PMOCs and POCs. Depicted is the representative H&E images and immunostaining pattern of CTSK, FGF1, GREM1, MMP13 and TIMP4 in POCs and PMOCs, respectively. (Original magnification: 200×).

Article Snippet: Serum concentrations of CTSK, MMP13, and FGF1 were measured by commercial, double-antibody, sandwich ELISA kits (Cusabio Life Science, Wuhan, China).

Techniques: Immunohistochemistry, Immunostaining

Journal: Journal of Cancer

Article Title: Identification of Cathepsin K in the Peritoneal Metastasis of Ovarian Carcinoma Using In-silico, Gene Expression Analysis

doi: 10.7150/jca.14277

Figure Lengend Snippet: Serum CTSK, CA125, MMP13, FGF1 and HE4 in ovarian tumors and normal controls*.

Article Snippet: Serum concentrations of CTSK, MMP13, and FGF1 were measured by commercial, double-antibody, sandwich ELISA kits (Cusabio Life Science, Wuhan, China).

Techniques: Control

Journal: Frontiers in Endocrinology

Article Title: Global Transcriptomic Analysis of the Canine corpus luteum (CL) During the First Half of Diestrus and Changes Induced by in vivo Inhibition of Prostaglandin Synthase 2 (PTGS2/COX2)

doi: 10.3389/fendo.2019.00715

Figure Lengend Snippet: List of gene symbols, corresponding gene names and TaqMan systems used for real time qPCR.

Article Snippet: FGF1 , Fibroblast growth factor 1 , XM_014108102.1 , Applied Biosystems, prod. no. Cf02716346_g1 , , 77.

Techniques: Sequencing, Derivative Assay

Journal: Frontiers in Endocrinology

Article Title: Global Transcriptomic Analysis of the Canine corpus luteum (CL) During the First Half of Diestrus and Changes Induced by in vivo Inhibition of Prostaglandin Synthase 2 (PTGS2/COX2)

doi: 10.3389/fendo.2019.00715

Figure Lengend Snippet: Relative gene expression of target candidate genes affected by time in control animals.

Article Snippet: FGF1 , Fibroblast growth factor 1 , XM_014108102.1 , Applied Biosystems, prod. no. Cf02716346_g1 , , 77.

Techniques: Gene Expression, Control