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: 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: 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: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Schematic of the experimental setup with THP1 or Raw264.7 cells with indicated treatment. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p b Quantification of proportion of TREM2 in THP1 and Raw264.7 cells with PTX. n = 3 biological independent samples. c Western blot of THP1 and Raw264.7 cells with indicated treatments. The experiment was independently repeated three times with similar results. d Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p e Quantification of proportion of TREM2 in THP1 incubated with CM of BT549 cells, SUM159 cells and MDA-MB-231 cells treated with PTX, in BMDM incubated with CM of Py8119 treated with PTX, in Raw264.7 incubated with CM from 4T1 and Py8119 cells treated with PTX, respectively. n = 3 biological independent samples. f Western blot analysis of TREM2 and related proteins in THP1, BMDMs, and Raw264.7 cells incubated with CM from BT549, SUM159, Py8119, and 4T1 cells, respectively. The experiment was independently repeated three times with similar results. g Representative immunofluorescence staining of THP1 cells incubated with BT549 CM. n = 3 biological independent samples. h Quantification of proportion of TREM2 in THP1 incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with Nab-PTX, respectively. n = 3 biological independent samples. i Quantification of the proportion of TREM2 in BMDM incubated with CM of Py8119 treated with Nab-PTX. n = 3 biological independent samples. j Cytokine array analysis of CM from BT549 cells treated indicated treatment. k Western blot analysis of the indicated proteins in Raw264.7 cells and BMDMs treated with recombinant FGF2. The experiment was independently repeated three times with similar results. l Quantification of the proportion of TREM2 in Raw264.7 cells and BMDMs treated with recombinant FGF2. n = 3 biological independent samples. m Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment, and then pretreatment using an FGF2 neutralizing antibody. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p n Western blot analysis of the indicated proteins in Raw264.7 and BMDMs incubated with the indicated CM. The experiment was independently repeated three times with similar results. o Quantification of the proportion of TREM2 in Raw264.7 and BMDMs incubated with the indicated CM. n = 3 biological independent samples. p Representative multiplex immunofluorescence staining of CD68, TREM2 and FGF2 in tumors treated with PTX or Nab-PTX ( n = 3 mice per group). The experiment was independently repeated three times with similar results. Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( b , e , h and i ) and two-sided one-way ANOVA followed by Tukey’s test ( l and o ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Recombinant, Multiplex Assay

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Overlap of RNA-seq ( n = 3) and PROMO public database analyses to predict transcription factors regulating TREM2 expression. b Correlations between FGF2 and ATF3 expression in breast cancer using TCGA databases. c qPCR analysis of Atf3 expression in the indicated cells. n = 4 (DMSO and PTX) or 3 (PBS and Nab-PTX) biological independent samples. d Western blot analysis of the indicated proteins in Py8119 cells transfected with ATF3-expressing or control vectors. The experiment was independently repeated three times with similar results. e qPCR analysis of Fgf2 expression in Py8119 cells transfected with Atf3 -expressing or control vectors. n = 3 biological independent samples. f Luciferase activity in HEK293T cells transfected with the indicated reporters and Atf3 -expressing or control vectors. n = 3 biological independent samples. g Abundance of Atf3 bound to the Trem2 promoter in Py8119 cells, as assessed by ChIP-qPCR. n = 3 biological independent samples. h ATAC-seq tracks showing the chromatin accessibility in the ATF3 loci for BT549 cells treated by PTX or Nab-PTX. n = 2 samples per group. Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( c, e and f ) and two-sided two-way ANOVA followed by Šídák’s test ( g ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: RNA Sequencing, Expressing, Western Blot, Transfection, Control, Luciferase, Activity Assay, ChIP-qPCR

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Overlap of GeneCards and EDCODE public databases analyses to predict transcription factors that regulate TREM2. b Western blot analysis of the indicated proteins in BMDMs and Raw264.7 cells incubated with FGF2 (left) and with CM from Py8119 cells treated with PTX (right). The experiment was independently repeated three times with similar results. c Western blot analysis of the indicated proteins in Raw264.7 cells incubated with the indicated treatment. The experiment was independently repeated three times with similar results. d qPCR analysis of Trem2 expression in BMDMs transfected with Egr1 -expressing vectors. n = 3 biological independent samples. e Western blot analysis of TREM2 expression in BMDMs transfected with Egr1 -expressing vectors. The experiment was independently repeated three times with similar results. f Luciferase activity of HEK293T cells transfected with the indicated reporters and EGR1 -expressing or control vectors. n = 3 biological independent samples. g Abundance of EGR1 bound to the TREM2 promoter in BMDMs assessed by ChIP-qPCR. n = 3 biological independent samples. h Schematic of the Transwell assay. The first CM was collected from tumor cells with indicated treatment, and the second CM was collected from macrophages incubated with the first CM. The migration and invasion capabilities of macrophages incubated with the first CM were assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p i Quantification of migration and invasion of Py8119 cells induced by BMDM CM incubated with CM from Py8119 cells treated with PTX, and BT549, SUM159, and MDA-MB-231 cells induced by THP1 CM incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with PTX (left) or Nab-PTX (right), respectively. n = 5 biological independent samples. j Cytokine array analysis of CM from BMDMs with or without TREM2. k Quantification of migration and invasion of Py8119 cells incubated with indicated proteins. n = 3 biological independent samples. l Quantification of migration and invasion of Py8119 cells induced by CM from BMDMs ( Trem2 +/+ ) with CDE-096. n = 3 biological independent samples. m Western blot analysis of EMT- stimulating proteins in BMDMs incubated with indicated proteins. The experiment was independently repeated three times with similar results. n Schematic illustration showing that FGF2 promotes ERK1/2 phosphorylation to upregulate EGR1, which increases TREM2 expression in macrophages. Upregulated TREM2 enhances the secretion of Serpin E1, HGF, CCL3, and CXCL2 from macrophages to tumor cells, facilitating tumor metastasis via EMT. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p . Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( d, f, i and l ), two-sided one-way ANOVA followed by Tukey’s test ( k ) and two-sided two-way ANOVA followed by Šídák’s test ( g ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: Western Blot, Incubation, Expressing, Transfection, Luciferase, Activity Assay, Control, ChIP-qPCR, Transwell Assay, Migration, Phospho-proteomics

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: PTX, but not Nab-PTX, promotes lung metastasis by inducing TREM2 + macrophage recruitment. Mechanistically, PTX enhances the ATF3-FGF2 axis in breast cancer cells; secreted FGF2 activates the EGR1–TREM2–EMT cytokine axis in macrophages. Created in BioRender. Xing, Y. (2026) https://BioRender.com/6hxlbow .

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques:

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: A Slco4c1 -/- mice were generated using Cas9/gRNA gene-editing techniques. Slco4c1 -/- mice were characterized by mouse tail genotyping ( B ), Western blot ( C ) and IF analyses ( D ). E The strategies for the establishment of Slco4c1 -/- and WT MASLD mice by feeding with GAN diet. F The percent of body weight (BW) changes in the GAN diet-fed Slco4c1 -/- mice ( n = 8 per time point) and WT mice ( n = 6 per time point). G The liver weights (LW) and the ratio of liver weights to body weights (LW/BW) in the GAN diet-fed Slco4c1 -/- mice ( n = 8) and WT mice ( n = 6). H WAT and BAT weights in the GAN diet-fed Slco4c1 -/- mice( n = 8) and WT mice ( n = 6). (I) serum ALT and AST levels in the GAN diet-fed Slco4c1 -/- mice( n = 8) and WT mice ( n = 6). J The levels of liver TG, FFA, and TC in the GAN diet-fed Slco4c1 -/- mice ( n = 8) and WT mice ( n = 6). K H&E (Scale bars:100 μm), Sirius red (Scale bars:100 μm), and Oil red O staining (Scale bars:50 μm) of liver sections from the GAN diet-fed WT ( n = 6) and Slco4c1 -/- ( n = 8) mice. L Liver histologic assessments of steatosis, lobular inflammation, hepatocyte ballooning, fibrosis and NAS score from the GAN diet-fed WT ( n = 6) and Slco4c1 -/- ( n = 8) mice. B: n = 3, biologically independent experiments, the results are similar. C , D An experiment was conducted on mice identified in Figure2B, yielding results consistent with those in Figure2B. F Plotted: mean ± SEM; Statistics: Multiple unpaired t test; *p < 0.05, ** p < 0.01, *** p < 0.001vs. the GAN diet-fed WT mice. G – J , L Each dot represents one mouse sample; Plotted: mean ± SD; Statistics: unpaired two-tailed Student’s t test; 95% confidence interval. *P < 0.05, ** p < 0.01, *** p < 0.001vs. the GAN diet-fed WT mice.

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Generated, Western Blot, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: A Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) of the lipidomic data from the livers of GAN diet-fed WT ( n = 6) and Slco4c1 -/- mice ( n = 6). B The lipid category ratios in mouse liver tissues. C The major components of liver lipid categories in the GAN diet-fed WT and Slco4c1 -/- mice. The relative levels of hepatic Fasn, Acc1, Scd1 mRNA transcripts ( D ) and protein expression ( E ) in the GAN diet-fed WT ( n = 6) and Slco4c1 -/- mice ( n = 8). F A diagram illustrates how SCLO4C1 inhibits lipid synthesis in the liver (Created with MedPeer (medpeer.cn). The relative levels of Fasn, Acc1, Scd1 mRNA transcripts ( G ) and protein expression ( H ) in primary mouse hepatocytes. The levels of TG ( I ) and TC ( J ) in primary mouse hepatocytes. K Oil red O staining in the BSA or PA-treated primary mouse hepatocytes from WT and Slco4c1 -/- mice. The relative levels of Fasn, Acc1, Scd1 mRNA transcripts ( L ) and protein expression ( M ) in different groups of HepG2 cells. The levels of TG ( N ) and TC ( O ) in different groups of HepG2 cells. P Oil red O staining of different groups of HepG2 cells. C The abscissa is the fold change after log2 conversion, and the ordinate is the lipid Sub Class. Each point in the figure represents a different lipid, the color of the point corresponds to different lipid Sub Class, and the size of the point is p after -log10 conversion. D , E Each dot represents one mouse sample; Plotted: mean ± SD; Statistics: unpaired two-tailed Student’s t test; 95% confidence interval. G n = 3 biologically independent experiments; Plotted: mean ± SD; Statistics: unpaired two-tailed Student’s t test; 95% confidence interval. H – P n = 3 biologically independent experiments; Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval. *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (PS Phosphatidylserine; PI Phosphatidylinositol; PG Phosphatidylglycerol; PE Phosphatidylethanolamine; PC Phosphatidylcholine; LPS Lyso-phosphatidylserine; LPG Lyso-phosphatidylglycerol; LPE Lyso-phosphatidylethanolamine; LPC Lyso-phosphatidylcholine; dMePE Dimethylphosphatidylethanolamine; SM Sphingomyelin; MGDG Monogalactosyldiacylglycerol; Cer Ceramides; DG Diglyceride).

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Expressing, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: A Luminescence Diagram of the “Persistently Open” Fluorescent Probe Jat-cAMP: The fluorophore was chemically conjugated to cAMP to form the Jat-cAMP probe, which was used for visualizing the transport of cAMP by SLCO4C1 protein. B Chemical synthesis of the Jat-cAMP probe. C The 2D highlighted the key interaction sites. D The 3D depiction illustrated the spatial binding conformation. E The strategies for determining the importance of SLCO4C1 Gln463 in transporting cAMP. F The CCK-8 analysis of the impact of Jat-cAMP on the viability of HepG2 cells. ( n = 6 Biological replicates; Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval.) G , H Live cell imaging of Jat-cAMP treated HepG2 cells. I ELISA analyses of cAMP concentrations in different groups of primary mouse hepatocytes. J ELISA analyses of cAMP concentrations in liver tissues of the GAN diet-fed WT( n = 6) and Slco4c1 -/- mice( n = 8). K ELISA analyses of cAMP concentrations in serum samples of the GAN diet-fed WT( n = 6) and Slco4c1 -/- mice( n = 8). L Correlation between hepatic Slco4c1 mRNA transcripts and cAMP levels in the GAN-fed WT mice. (Person r, two-tailed, 95% confidence interval) ( M ) Relative levels of PKAc and p-Creb in different groups of primary mouse hepatocytes. N Relative Srebp1 mRNA transcripts in different groups of primary mouse hepatocytes. ( n = 6 Biological replicates; Plotted: mean ± SD; Statistics: unpaired two-tailed Student’s t test; 95% confidence interval.) O Relative Srebp1c protein levels in different groups of primary mouse hepatocytes. P Relative Epac1 protein levels in the different groups of primary mouse hepatocytes. H , I n = 5 Biological replicates; Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval. J-K: Each dot represents one mouse sample; Plotted: mean ± SD; Statistics: unpaired two-tailed Student’s t test; 95% confidence interval. *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. M , O , P n = 3 per group, Biological replicates; Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval. * p < 0.05 vs. the BSA-treated cells from WT mice; # p < 0.05 vs. the PA-treated cells from WT mice. $ p < 0.05 vs. the BSA-treated cells from Slco4c1 -/- mice.

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Binding Assay, CCK-8 Assay, Live Cell Imaging, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: A Cytokines stimulated Slco4c1 mRNA transcription in primary mouse hepatocytes ( B ) FGF21 expression levels in different hepatic cell types. C Hepatic Fgf21 mRNA levels in GAN diet fed mice. D Correlation analysis between hepatic Fgf21 and Slco4c1 mRNA levels ( n = 6). E , F Treatment with FGF21 stimulated Slco4c1 mRNA and protein expression in primary mouse hepatocytes ( G ) Schematic illustration of the predicted transcription factors. H The predicted transcription factor mRNA transcripts in the FGF21-treated primary mouse hepatocytes. I , J Hepatic Egr1/2/3 mRNA transcripts and protein expression in the indicated groups of mice. (* p < 0.05, vs. Chow diet-WT) ( K ) Correlation between hepatic Slco4c1 and Egrs mRNA levels in the GAN diet-fed WT mice ( n = 6). L A diagram illustrating the potential EGRs binding sites in human SLCO4C1 proximal promoter and the reporter constructs. M Luciferase activities in 293 T cells co-transfected with truncated SLCO4C1 promoters with, or without, plasmids for EGRs overexpression. N A schematic diagram displayed the EGR1-binding site in the SLCO4C1 promoter and its mutant. O Luciferase activity in 293 T cells co-transfected with the mutant Slco4c1 promoter and the plasmid for EGR1 overexpression. Relative levels of Egr1 mRNA ( P ) and protein ( Q ) expression in primary mouse hepatocytes treated with, or without, an ERK/MAPK inhibitor, and after stimulated by FGF21 were examined. R ChIP assay revealed that FGF21 increased Egr1 binding to the Slco4c1 -59 promoters in primary mouse hepatocytes. Relative levels Slco4c1 mRNA (S) and protein (T) expression in primary mouse hepatocytes treated with, or without, an ERK/MAPK inhibitor, and after stimulated by FGF21 were examined. A n = 5 and M , O n = 3, Biological replicates; C , I , J Each dot represents one mouse sample; Statistics: unpaired two-tailed Student’s t test. R: n = 3 and E , F , H , P , Q , S , T n = 5, Biological replicates; Statistics: one-way ANOVA test, Multiple comparisons. *p < 0.05, * *p < 0.01, *** p < 0.001, **** p < 0.0001. F: * p < 0.05 vs. FGF21 0 μg/ml; # p < 0.05 vs. FGF21 0.1 μg/ml. Q, T: * p < 0.05 vs. FGF21 0 μg/ml; # p < 0.05 vs. FGF21 1 μg/ml. All data statistics are presented as mean ± SD and 95% confidence interval.

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Expressing, Binding Assay, Construct, Luciferase, Transfection, Over Expression, Mutagenesis, Activity Assay, Plasmid Preparation, Two Tailed Test

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: A Schematic illustrating of the experiment design of the GAN diet-induced MASL and MASH mouse models. B Bioluminescence images displayed the AAV8-mediated Slco4c1 expression in mice 12 weeks after inoculation. C The dynamic changes in body weights in the different groups of mice. (Plotted: mean ± SEM; Statistics: two-way ANOVA test, Multiple comparisons, 95% confidence interval.) D Liver weights and liver weight/body weight ratios (LW/BW) of MASL and MASH mice. E White and brown adipose tissue weights of MASL and MASH mice. H&E, Sirius red and Oil red O staining of liver tissue sections of MASL F and MASH ( G ) mice. Quantitative liver histologic assessment of the severity of MASL ( H ) and MASH ( I ) mice, including steatosis, lobular inflammation, hepatocyte ballooning, fibrosis and NAS score. The levels of serum ALT, AST ( J ) and TG, TC ( K ) in MASL and MASH mice. L Mice were injected with control virions or AAV8-TBG-Luciferase- Slco4c1 and their serum and hepatic cAMP levels were quantified using ELISA. M , N Western blot revealed that induction of Slco4c1 overexpression by injection with AAV8-TBG-Luciferase- Slco4c1 increased the relative levels of PKAc and p-Creb(S133) proteins ( M ), but decreased the relative levels of Srebp1 , Fasn , Acc1 , and Scd1 mRNA transcripts in the liver of mice, compared with that in the control mice ( N ). C – E , H – K n = 4 per group in MASL model. In MASH model, Chow diet group and GAN diet-AAV8-TBG-Luciferase-CTR group n = 5, AAV8-TBG-Luciferase- Slco4c1 group n = 6; D , E , H – K Each dot represents one mouse sample, Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval. *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. L , N Chow diet group and GAN diet-AAV8-TBG-Luciferase-CTR group n = 9, AAV8-TBG-Luciferase- Slco4c1 group n = 10, each dot represents one mouse sample, Plotted: mean ± SD; Statistics: one-way ANOVA test, Multiple comparisons, 95% confidence interval. *p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. M: * P < 0.05 vs. the Chow diet group, # P < 0.05 vs. the GAN diet-AAV8-TBG-Luciferase-CTR group.

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Expressing, Staining, Injection, Control, Luciferase, Enzyme-linked Immunosorbent Assay, Western Blot, Over Expression

Journal: Nature Communications

Article Title: Hepatocyte SLCO4C1 is a cAMP uptake transporter for inhibiting lipogenesis and a therapeutic target for MASLD

doi: 10.1038/s41467-026-70729-0

Figure Lengend Snippet: During the pathogenic process, metabolic disorder stimulates FGF21 expression, which through its receptors, activates the ERK/MAPK signaling to induce EGR1 expression. The transcription factor of EGR1 binds to the SLCO4C1 promoter and up-regulates its expression in hepatocytes. The SLCO4C1 functions as a cAMP uptake transporter through the interaction of its Gln463 with cAMP, increasing intracellular cAMP levels that activate the PKA-CREB signaling to feedback down-regulate SREBP1 and downstream ACC1, FASN and SCD1 expression, inhibiting fatty acid synthesis. Accordingly, induction of SLCO4C1 overexpression by AAV8-mediated hepatic SLCO4C1 expression and/or increasing intracellular cAMP levels by the Forskolin treatment effectively prevent and mitigate the progression of MASLD. Therefore, SLCO4C1 is a therapeutic target for MASLD. Conceivably, our findings may aid in the design of therapeutic strategies for treating MASLD.

Article Snippet: Primary mouse hepatocytes were treated with FGF21 (1 μg/ml) or first treated with an inhibitor of the ERK/MAPK signaling (PD98059, 25 μM, MCE, New Jersey, USA, Cat: HY-12028) for 1 h and subsequently treated with FGF21.

Techniques: Expressing, Over Expression

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: 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