Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Comparison of serum Vit B 6 and serological metabolism parameters among control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Comparison, Control

Journal: BMJ Open

Article Title: Association between serum vitamin B 6 concentration and risk of osteoporosis in the middle-aged and older people in China: a cross-sectional study

doi: 10.1136/bmjopen-2018-028129

Figure Lengend Snippet: Relationship between serum Vit B 6 concentrations and bone metabolism parameter concentrations in control, osteopenia and osteoporosis groups

Article Snippet: The concentration of fibroblast growth factor 23 (FGF23) was measured through an ELISA performed using a human FGF23 ELISA kit (CSB-E10113h, Cusabio Biotech, Wuhan, China) and a micro plate reader (MK3, Thermo, Waltham, Massachusetts, USA).

Techniques: Control

Journal: Nature Communications

Article Title: Transgenerational inheritance of diminished ovarian reserve triggered by prenatal propylparaben exposure in mice

doi: 10.1038/s41467-025-63440-z

Figure Lengend Snippet: a STRING database revealed strong interactions between RhoBTB1 and FGF18, RhoBTB1 and CTSK, respectively. b , c The protein levels of FGF18 and MAPK signaling pathway in KGN cells detected by western blotting following PrP exposure. Data from ( c ) are representative of three independent experiments (mean ± SEM; one-way ANOVA). d , e The protein levels of FGF18 and MAPK signaling pathway in KGN cells detected by western blotting following RhoBTB1 over-expression. Data from ( e ) are representative of three independent experiments (mean ± SEM; unpaired two-tailed t -test). f , g The protein levels of FGF18, RhoBTB1, and MAPK signaling pathway in GCs detected by western blotting following PrP exposure and siRNA-F02 interference of Fgf18 . Data from ( g ) are representative of three independent experiments (mean ± SEM; one-way ANOVA). h , i The protein levels of RhoBTB1, FGF18, and MAPK signaling pathway in GCs detected by western blotting following PrP exposure and siRNA-R03 interference of Rhobtb1 . Data from ( i ) are representative of three independent experiments (mean ± SEM; one-way ANOVA). j , k The protein levels of FGF18 in GCs following PrP exposure and MLN4924 treatment. Data from ( k ) are representative of three independent experiments (mean ± SEM; one-way ANOVA). l , m The protein levels of FGF18 in GCs following RhoBTB1 over-expression and MLN4924 treatment. Data from ( m ) are representative of three independent experiments (mean ± SEM; unpaired two-tailed t -test). n , o The protein levels of FGF18 in GCs following PrP exposure and MG132 treatment. Data from ( o ) are representative of three independent experiments (mean ± SEM; one-way ANOVA). p , q The protein levels of FGF18 in GCs following RhoBTB1 over-expression and MG132 treatment. Data from ( q ) are representative of three independent experiments (mean ± SEM; unpaired two-tailed t -test). The samples derived from the same experiment and that blots were processed in parallel. Source data are provided as a Source Data file.

Article Snippet: The Antibodies were purchased from different companies: Cell Signaling Technology for ERK (Extracellular Signal-Regulated Kinase; 4695T), p-ERK (Phosphorylated ERK; 4370T), p38 (p38 Mitogen-Activated Protein Kinase; 8690T), p-p38 (Phosphorylated p38; 4511T), JNK (c-Jun N-terminal Kinase; 9252T), and p-JNK (Phosphorylated JNK; 4668T); Abclonal for β-ACTIN (AC038), active-Caspase3 (A19654), BCL-2 (A19693), BAX (A12009), SOD2 (A19576), BMP15 (A7321), CAT (A11777), HSD17B7 (A17157), and CTSK (A1782); Proteintech for FGF18 (60341-1-lg); Immunoway for RhoBTB1 (YT4084); Servicebio for HRP conjugated goat anti-rabbit IgG (GB23303), and Alexa Fluor® 488-conjugated goat anti-rabbit IgG (GB25303).

Techniques: Western Blot, Over Expression, Two Tailed Test, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Construction, purification, and characterization of FGF21-164. ( A ) FGF21-164 sequence and FGF21 sequence. In the FGF21-164 sequence, red marked the mutated amino acid and blue represents the TR acid sequence from hCD164. In the FGF21 sequence, the red-labeled amino acid represents the amino acid sequence whose n-terminal is deleted in FGF21-164. ( B ) Subcloned cell culture supernatants expressing FGF21-164 and FGF21 were analyzed using Western blotting (Lane M: molecular weight marker. Lane 1~7: FGF21-164 clones 1, 2, 3, A4, B2, F3, and H4. Line 8: FGF21 clone2). ( C ) The purified FGF21-164 and FGF21 were assayed using SDS-PAGE and Western blotting (Lane M: molecular weight marker. Lane 1: purified FGF21-164. Line 2: purified FGF21). ( D ) Purity and hydrodynamic radius of FGF21-164 were analyzed using SEC-HPLC. ( E ) MALDI-TOF mass-spectrometry analysis of FGF21-164. ([M + H] + denotes the singly charged ionic forms, and [M + 2H] 2+ denotes doubly charged ionic forms). ( F ) Circular Dichroism (CD) spectra of FGF21-164 and FGF21.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Purification, Sequencing, Labeling, Cell Culture, Expressing, Western Blot, Molecular Weight, Marker, Clone Assay, SDS Page, Mass Spectrometry, Circular Dichroism

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Glycan forms analysis of the protein FGF21-164. ( A ) O-linked glycan forms of FGF21-164. ( B ) N-linked glycan forms of FGF21-164.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of FGF21-164 in adipocytes induced by 3T3-L1. ( A ) The process of inducing 3T3-L1’s differentiation into adipocytes. ( B ) Reduction in glucose in medium stimulated by FGF21-164 in adipocytes induced by 3T3-L1. Data are means ± SEM (n = 6). * p < 0.05, *** p < 0.001, and **** p < 0.0001 compared to 0 μg/mL FGF21-164. ( C ) Phospho-Erk1/2-specific bands (Thr202/Tyr204; 44–42 kDa) were detected in 3T3-L1-derived adipocytes after FGF21-164 stimulation. ( D ) Evaluation of lipid droplet accumulation in adipocytes induced by 3T3-L1 with or without FGF21-164. The scale bar represents 50 μm.Data are the means ± SEM (n = 6). * p < 0.05, **** p < 0.0001.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Pharmacokinetics of FGF21-164. FGF21-164 plasma levels in C57BL/6 mice as determined via targeted LC-MS after i.v. (tail vein) bolus injection of FGF21-164 protein.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Liquid Chromatography with Mass Spectroscopy, Injection

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: The results of an oral glucose tolerance test applied to ob / ob mice after a single FGF21-164 treatment. ( A ) Blood glucose levels were measured at 0, 2, 3, 4, 5, 6, 7, and 8 h after the administration of FGF21-164. Data are the means ± SEM (n = 4–5). * p < 0.05 and ** p < 0.01 compared with the model. ( B ) The decrease in blood glucose levels post-administration relative to the baseline measurement at 0 h. Data are the means ± SEM (n = 4–5). * p < 0.05 compared with the model.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of a single administration of FGF21-164 on ob / ob mice. ( A , D , G ) Fasting glucose levels. Blood glucose levels were measured at 7, 21, and 28 days after a single dose of FGF21-164 was administered. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( B , E , H ) The OGTT at 7, 21, and 28 days. After oral glucose administration, blood glucose levels were measured. Data are the means ± SEM (n = 4–5). * p < 0.05, *** p < 0.001, and **** p < 0.0001 at 6 mg·kg −1 vs. model. # p < 0.05, ## p < 0.01 and #### p < 0.0001 at 12 mg·kg −1 vs. model. ( C , F , I ) The glucose AUC during the OGTT process. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, *** p < 0.001. ( J ) Body weight measured at 0, 7, 21, and 28 days. Data are the means ± SEM (n = 4–5). ns: no significance.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of repeated administration of FGF21-164. ( A ) Body weight changes. The DIO mice were treated with PBS or FGF21-164 once every 2 days. Data are the means ± SEM (n = 5–7). **** p < 0.0001 vs. Ctrl. #### p < 0.0001 for FGF21-164 vs. the model. ( B ) The liver sections were stained with Oil-red-O. The scale bar represents 100 μm. DIO mice showed many red lipid droplets in hepatic tissue (indicated by the black arrow), while lipid droplet levels were lower after the FGF21-164 treatment.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Staining

Journal: bioRxiv

Article Title: KGF induces podosome formation via integrin-Erk1/2 signaling in human immortalized oral epithelial cells

doi: 10.1101/508416

Figure Lengend Snippet: KGF Mediated its Effects on HIOECs through Erk1/2 and Akt Signaling. (A and B) Western blot detection of the expression level of KGFR in HIOECs after KGF treatment, (C and D) the phosphorylation level of Jak2 and Stat3 after KGF stimulation for 15 min, and (E and F) the phosphorylation level of Erk1/2, p38, Jnk, and Akt after KGF stimulation for 15 min. n=3 independent experiments. Analysis of matrix protein expression in protein extracts from each group by ImageJ. (G) EdU labeling assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. (H) The quantification shown in the right graph indicates the mean ± SEM. (I) MTT assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. Data are expressed as the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: HIOECs at 3x10 4 cells/well were seeded in the 96-well plate and exposed to KGF (10 ng/mL; Proteintech) for 24 h. For MTT assays, the cells were incubated with 20 μL of MTT solution (5 mg/mL) at 37°C for 4 h. The supernatant was discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well.

Techniques: Western Blot, Expressing, Phospho-proteomics, Labeling, MTT Assay

Journal: International Journal of Biological Sciences

Article Title: Generation of VEGF knock-in Cashmere goat via the CRISPR/Cas9 system

doi: 10.7150/ijbs.55559

Figure Lengend Snippet: Production of Knock-in cell colony. A. Gene-targeting strategy of Cas9-mediated knock-in of VEGF or DsRed2 to the FGF5 locus. B. Gene names, types, and target sequences of the top 10 potential off-target sites of sgRNA were used in this study. C. Process of exploring the optimal HDR efficiency of small molecules. D. Spanning the homologous arms PCR identification results of cell colonies. E. Images of cell colonies at 3, 7, and 15 days. Scale bar = 100 µm. F. The sequences and locations of target sites at exon 1 of FGF5 gene.

Article Snippet: Tris Buffered (CWBIO, Beijing, China) saline Tween was used to wash the nitrocellulose filter membrane for 5 min. After blocking the membranes with 5% skim milk (Difco TM Skim Milk, BD Biosciences) for 2 h, alpha-tubulin antibody (Proteintech, Wuhan, China), FGF5 antibody (Proteintech, Wuhan, China), and VEGF antibody (Proteintech) were added at a dilution of 1:1000 in 5% bovine serum albumin, and the samples incubated at 4 °C overnight.

Techniques: Knock-In

Journal: International Journal of Biological Sciences

Article Title: Generation of VEGF knock-in Cashmere goat via the CRISPR/Cas9 system

doi: 10.7150/ijbs.55559

Figure Lengend Snippet: VEGF knock-in Cashmere goats and its identification and detection. A. Images of 1-month-old SCNT and GEC. B. Images of 15-month-old SCNT, GEC, and WT. Red arrows indicate tufts of cashmere. C. Spanning the homologous arms PCR identification results of WT, SCNT and GEC. D. Southern blot identification results of SCNT, GEC, and WT. E. VEGF and FGF5 expression in skin tissues of WT, SCNT and GEC by Western blot. F, G. RT-PCR was used to detect the expression of VEGF and FGF5 in skin tissues of S WT, SCNT and GEC. H. Routine blood tests were performed on WT, SCNT and GEC, and the number of WBC was counted.

Article Snippet: Tris Buffered (CWBIO, Beijing, China) saline Tween was used to wash the nitrocellulose filter membrane for 5 min. After blocking the membranes with 5% skim milk (Difco TM Skim Milk, BD Biosciences) for 2 h, alpha-tubulin antibody (Proteintech, Wuhan, China), FGF5 antibody (Proteintech, Wuhan, China), and VEGF antibody (Proteintech) were added at a dilution of 1:1000 in 5% bovine serum albumin, and the samples incubated at 4 °C overnight.

Techniques: Knock-In, Southern Blot, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Digital Gene Expression Tag Profiling Analysis of the Gene Expression Patterns Regulating the Early Stage of Mouse Spermatogenesis

doi: 10.1371/journal.pone.0058680

Figure Lengend Snippet: (A) Protein levels of FGF13, SFRP2 and RAC in GC-1spg and GC-2spd (ts) cells were detected by western blotting respectively. β-Actin were used to normalize the individual expression levels. (B) Protein level of WNT10A and FGF7 in the medium supernatants from GC-1spg and GC-2spd (ts) cells were determined by ELISA. The data are presented as the mean plus SEM from a representative of three independent experiments, with each performed in triplicate. Statistically significant differences between GC-1spg and GC-2spd (ts) cells are indicated above the bars: * means P<0.05.

Article Snippet: The antibodies used in Western blotting were RAC (ab13048, Abcam, USA), SFRP2 (12189-1-AP, Proteintech, USA), FGF13 (13201-1-AP, Proteintech,USA) and β-actin (A-4700, Sigma, USA).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay