Journal: World Journal of Gastroenterology

Article Title: FBP1 as a key regulator of focal adhesion kinase-mediated hepatic stellate cell activation: Multi-omics and experimental validation

doi: 10.3748/wjg.v31.i28.107361

Figure Lengend Snippet: Transcriptomics analysis of differential gene expression. A: Principal component analysis (PCA) of normalized counts PCA showing gene expression variance among normal, carbon tetrachloride (CCl 4 ), and PF562271 (PF) conditions. The first principal component (PC1) explains 59% of the variance, and PC2 explains 8%; B: Volcano plot showing differentially expressed genes (DEGs) between the normal and CCl 4 groups. Significant genes (log2 fold change > 1 or < -1, P < 0.05) are highlighted; C: Volcano plot comparing gene expression between the CCl 4 and PF groups. Significant genes are marked similarly; D: Mfuzz clustering of gene expression. Six gene clusters showing expression changes across the normal, CCl 4 , and PF groups, highlighting condition-specific gene regulation; E: UpSet plot showing the overlap of DEGs between comparisons; F: Venn diagram showing unique and shared DEGs between the normal vs CCl 4 and CCl 4 vs PF groups.

Article Snippet: Recombinant FBP1 protein (HY- P70275 ) and PF562271 were obtained from MedChemExpress.

Techniques: Gene Expression, Expressing

Journal: World Journal of Gastroenterology

Article Title: FBP1 as a key regulator of focal adhesion kinase-mediated hepatic stellate cell activation: Multi-omics and experimental validation

doi: 10.3748/wjg.v31.i28.107361

Figure Lengend Snippet: Proteomics analysis of differential protein expression. A: Principal component analysis (PCA) of normalized protein counts PCA of protein expression across normal, carbon tetrachloride (CCl₄) and PF562271 (PF) conditions. The first principal component (PC1) explains 39.3% of variance, while PC2 explains 9.5%. The three conditions show clear separation; B: Volcano plot showing differentially expressed proteins between the normal and CCl₄ groups. Significant proteins ( P < 0.05, log2 fold change > 1 or < -1) are highlighted; C: Volcano plot comparing the CCl₄ and PF groups. Significant proteins are marked, indicating differential expression between these two conditions ( P < 0.05); D: Mfuzz clustering of protein expression profiles. Clusters 1, 3, and 4 are shown, highlighting distinct expression trends in normal, CCl₄, and PF conditions; E: UpSet plot showing overlaps of differentially expressed proteins between comparisons (normal vs CCl₄, CCl₄ vs PF) and clusters from Mfuzz analysis; F: Venn diagram showing overlap of differentially expressed proteins between the normal vs CCl₄ and CCl₄ vs PF groups. The diagram indicates unique and shared proteins across the conditions.

Article Snippet: Recombinant FBP1 protein (HY- P70275 ) and PF562271 were obtained from MedChemExpress.

Techniques: Expressing, Quantitative Proteomics

Journal: Biology

Article Title: KDM6B Variants May Contribute to the Pathophysiology of Human Cerebral Folate Deficiency

doi: 10.3390/biology12010074

Figure Lengend Snippet: Overexpression of KDM6B mutants affected FOLR1 protein level and folate uptake ability of HeLa cells. ( A ) HeLa cells were transfected with mutated and wild-type constructs of Flag-tagged KDM6B and pcDNA3.1 backbone vector for 48 h, and Western blotting was performed to quantify FOLR1 protein level in each group. GAPDH was used as loading control. ( B ) Western blotting was repeated three times and a Student t -test was performed to compare the protein levels between wild-type and mutant. Significance compared to WT: “a” represents p < 0.001; “c” represents p < 0.05.

Article Snippet: 48 h after transfection, cells were fixed with 4% paraformaldehyde for 30 min and cell membranns were permeabilized with 0.3% Triton X-100 in TBST for 20 min, and then blocked with 10% normal goat serum (NGS) in Tris-buffered saline with 0.1% Tween ® 20 Detergent (TBST) for 1 h. Cells were then incubated with FOLR1 antibody (1:100, Proteintech, Rosemont, IL, USA, 23355-1-AP) and flag antibody (1:1000, ThermoFisher Scientific, MA1-91878) overnight at 4 °C.

Techniques: Over Expression, Transfection, Construct, Plasmid Preparation, Western Blot, Control, Mutagenesis

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 2 | FBP1 and PCK1 undergo Kbhb in a time‐ and dose‐dependent manner. (A) HEK293T cells transfected with Flag‐FBP1, Flag‐FBP2, and Flag‐PCK1 were treated with 10‐mM BHB for 24 h and Kbhb modification was detected. (B, C) Kbhb modification of Flag‐FBP1 (B) and Flag‐ PCK1 (C) in HEK293T cells treated with 10‐mM BHB for the indicated time. (D, E) Kbhb modification of Flag‐FBP1 (D) and Flag‐PCK1 (E) in HEK293T cells treated with indicated concentrations of BHB for 24 h.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Transfection, Modification

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 3 | BHB enhances glucose levels in bovine hepatocytes. (A) The bovine hepatocytes were treated with the indicated concentration of BHB, and the glucose concentrations were detected by kit. (B–D) The bovine hepatocytes were treated with indicated concentration of BHB, and endogenous expression of FBP1, PCK1, and G6PC genes was examined by RT‐qPCR. (E) The bovine hepatocytes were treated with indicated concentration of BHB, and the indicated proteins were detected by Western blot. (F–H) Quantitative data for the indicated protein level was presented. Statistical analyses were performed with the Student's t test and one‐way ANOVA. *p < 0.05, ***p < 0.001, ns stands for no significant change.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Concentration Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 4 | BHB is transported into the cell via MCT1. (A, B) Kbhb modification of Flag‐FBP1 (A) and Flag‐PCK1 (B) in HEK293T cells treated with 10‐mM BHB in combination with 10‐μM AZD3965 for 24 h. (C, D) MCT1 knockdown by siRNA transfection. Flag‐FBP1 (C) and Flag‐PCK1 (D) Kbhb levels were analyzed by immunoblotting with indicated antibodies. (E) The bovine hepatocytes were treated with BHB and 10‐μM AZD3965, and the glucose concentrations were detected by kit. (F) The bovine hepatocytes were treated with BHB and MCT1 knockdown by siRNA, and the glucose concentrations were detected by kit. Statistical analyses were performed with the one‐way ANOVA. ***p < 0.001.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Modification, Knockdown, Transfection, Western Blot

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 5 | Kbhb of FBP1 and PCK1 is catalyzed by p300 and removed by HDAC. (A, B) P300 knockdown by siRNA in HEK293T cells. Flag‐FBP1 (A) and Flag‐PCK1 (B) Kbhb levels were analyzed by immunoblotting with indicated antibodies. (C, D) Immunoprecipitated FBP1 (C) and PCK1 (D) proteins were incubated with recombinant p300 protein and β‐hydroxybutyryl‐CoA (BHB‐CoA, 20 μM) for 1 h at 30°C, pH 8.0. (E, F) Kbhb modification of Flag‐FBP1 (E) and Flag‐PCK1 (F) in HEK293T cells treated with 10‐mM BHB in combination with NAM or TSA for 24 h. (G) The bovine hepatocytes with p300 knockdown were treated with 10‐mM BHB for 24 h and the glucose concentrations were detected by kit. (H) The bovine hepatocytes were treated with BHB and 50‐nM TSA, and the glucose concentrations were detected by kit. Statistical analyses were performed with the one‐way ANOVA. ***p < 0.001.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Knockdown, Western Blot, Immunoprecipitation, Incubation, Recombinant, Modification

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 6 | Kbhb of FBP1 and PCK1 promotes their activity. (A) The bovine hepatocytes were treated with 10‐mM BHB and CHX for the indicated time, and then Western blot was used to evaluate the levels of the indicated proteins. (B, C) The bovine hepatocytes were treated with 10‐mM BHB alone or in combination with 10‐μM AZD3965 for 24 h, and the activity of FBP1 (B) and PCK1 (C) was detected by ELISA kit. (D, E) The bovine hepatocytes were treated with 10‐mM BHB alone or in combination with MCT1 knockdown by siRNA, and the activity of FBP1 (D) and PCK1 (E) was detected by ELISA kit. (F, G) The bovine hepatocytes were treated with 10‐mM BHB alone or in combination with p300 knockdown by siRNA, and the activity of FBP1 (F) and PCK1 (G) was detected by ELISA kit. Statistical analyses were performed with the one‐ way ANOVA. ***p < 0.001.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 7 | FBP1 and PCK1 Kbhb occur at lysine 43 and 191, respectively. (A) Schematic representation of experimental workflow for the identification of Kbhb‐containing protein substrates in HEK293T cells. (B, C) Illustration of PCK1 K191bhb (B) and FBP1 K43bhb (C) identified by mass spectrometry (MS). (D) Detecting FBP1 Kbhb in HEK293T cells transfected with FBP1 WT and K34R mutant plasmids and treated with 10‐mM BHB for 24 h. (E) Detecting PCK1 Kbhb in HEK293T cells transfected with PCK1 WT and K191R mutant plasmids and treated with 10‐ mM BHB for 24 h. (F, G) Immunoprecipitated FBP1 WT and K43R (F) and PCK1 WT and K191R (G) mutant plasmids were incubated with recombinant p300 protein and BHB‐CoA for 1 h at 30°C, pH 8.0, and the levels of Kbhb were detected by Western blot.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Mass Spectrometry, Transfection, Mutagenesis, Immunoprecipitation, Incubation, Recombinant, Western Blot

Journal: Animal Research and One Health

Article Title: Compensation Response to Hepatic Gluconeogenesis via β‐Hydroxybutyrylation of FBP1 and PCK1 in Dairy Cows

doi: 10.1002/aro2.70022

Figure Lengend Snippet: FIGURE 8 | BHB increases glucose levels and activity of FBP1 and PCK1 through K43bhb and K191bhb, respectively. (A, B) The bovine hepatocytes transfected with FBP1 WT or K43R and treated with indicated concentrations of BHB were utilized for glucose concentration (A) and Western blot (B). (C, D) The bovine hepatocytes transfected with PCK1 WT or K191R and treated with indicated concentrations of BHB were utilized for glucose concentration (C) and Western blot (D). (E, F) The bovine hepatocytes transfected with FBP1 WT or K43R and treated with indicated concentrations of BHB were utilized for the activity of FBP1 (E) and Western blot (F). (G, H) The bovine hepatocytes transfected with PCK1 WT or K191R and treated with indicated concentrations of BHB were utilized for the activity of PCK1 (G) and Western blot (H). (I, J) HEK293T cells transfected with Flag‐FBP1 and Flag‐PCK1 from humans and cows were treated with indicated concentrations of BHB for 24 h, and Kbhb modification was detected. Statistical analyses were performed with the Student's t test. *p < 0.05, ***p < 0.001, ns stands for no significant change.

Article Snippet: FBP1 (12842‐ 1‐AP), PCK1 (16754‐1‐AP), G6PC (66860‐1‐Ig), and β‐actin (66009‐1‐Ig) antibodies were obtained from Proteintech (Chicago, USA).

Techniques: Activity Assay, Transfection, Concentration Assay, Western Blot, Modification

Journal: BMC Oral Health

Article Title: Folate-receptor 1 level in periodontal disease: a pilot study

doi: 10.1186/s12903-019-0909-z

Figure Lengend Snippet: The Spearman’s rank correlation (r) among clinical parameters and FOLR1 levels

Article Snippet: FOLR1 levels were assayed by Human FOLR1 kit (Boster, CA, USA) using enzyme-linked immune sorbent assay (ELISA) method.

Techniques:

Journal: Cell systems

Article Title: Four key steps control glycolytic flux in mammalian cells

doi: 10.1016/j.cels.2018.06.003

Figure Lengend Snippet:

Article Snippet: FBP1: FBP1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_000507","term_id":"1677501380","term_text":"NM_000507"}} NM_000507 ) Human cDNA Clone , Origene , Cat# SC119828.

Techniques: Recombinant, Modification, Transfection, Clone Assay, Enzyme-linked Immunosorbent Assay, Colorimetric Assay, Lactate Dehydrogenase Assay, Bicinchoninic Acid Protein Assay, PK Assay, Plasmid Preparation, Sequencing, Expressing, Variant Assay, Cell Culture