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: 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: Cell metabolism

Article Title: Hyodeoxycholic acid alleviates non-alcoholic fatty liver disease through modulating the gut-liver axis.

doi: 10.1016/j.cmet.2023.07.011

Figure Lengend Snippet: Figure 4. HDCA alleviated hepatic steatosis in a gut-dependent manner (A) The mRNA relative expression of intestinal FXR signaling-related genes (Fxr and Fgf15) from experiment 4 (5 biological replicates for each group). (B) The protein expression of intestinal FGF15 (n = 3, representative of 3 biological replicates for each group).

Article Snippet: For the FGF15 measurement, the level of serum FGF15 was detected using a fibroblast growth factor 15 ELISA kit (CSB-EL522052MO, CUSABIO).

Techniques: Expressing

Journal: Cell metabolism

Article Title: Hyodeoxycholic acid alleviates non-alcoholic fatty liver disease through modulating the gut-liver axis.

doi: 10.1016/j.cmet.2023.07.011

Figure Lengend Snippet: Figure 7. The molecular mechanism of amelioration of hepatic steatosis by HDCA HDCA ameliorated hepatic steatosis by shifting the BA classic (downregulated CYP7A1) and alternative (upregulated CYP7B1) synthesis pathways. Two mechanisms initiated by HDCA worked simulta- neously: (1) HDCA inhibited intestinal FXR-FGF15/ 19 signaling, thus activating both CYP7A1 and CYP7B1, and (2) HDCA remodeled the structure of gut microbiota, enriching the P. distasonis and its fatty acid metabolite, g-linolenic acid. g-Linolenic acid inhibited the hepatic CYP7A1 through PPARa signaling.

Article Snippet: For the FGF15 measurement, the level of serum FGF15 was detected using a fibroblast growth factor 15 ELISA kit (CSB-EL522052MO, CUSABIO).

Techniques:

Journal: Journal of Orthopaedic Surgery and Research

Article Title: GLP-2 alleviates postmenopausal osteoporosis by acting on osteoblasts through the PI3K/AKT/FOXO1 signalling pathway to downregulate FGF23 expression

doi: 10.1186/s13018-026-06661-2

Figure Lengend Snippet: Transcriptomic analysis of femoral tissues from sham, OVX, and OVX + GLP-2 groups. A Scatter plot of DEGs between the OVX and sham groups (1654 upregulated genes and 2038 downregulated genes). B Scatter plot of DEGs between the OVX + GLP-2 and OVX groups (318 upregulated genes and 396 downregulated genes). C Heatmap of 24 representative DEGs among the three groups. D GO enrichmentresults showing DEGs involved in cellular component, biological process, and molecular function categories. E KEGG signalling pathway diagram. F KEGG enrichment bar chart. G KEGG enrichment scatter plot. H Relative mRNA expression of FGF23, FGF6, and FGF18 in the tibiae of the OVX group

Article Snippet: FGF23 levels in cell culture supernatants were measured using a mouse FGF23 ELISA Kit (CSB-EL008629MO, Cusabio) following the manufacturer’s protocol.

Techniques: Expressing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: GLP-2 alleviates postmenopausal osteoporosis by acting on osteoblasts through the PI3K/AKT/FOXO1 signalling pathway to downregulate FGF23 expression

doi: 10.1186/s13018-026-06661-2

Figure Lengend Snippet: Effects of GLP-2 and the PI3K inhibitor LY294002 on FGF23 expression in OVX mice. Forty-four female C57BL/6J mice were randomly divided into five groups: sham-operated (sham, n = 6), OVX treated with vehicle (OVX, n = 8), OVX treated with GLP-2 (OVX + GLP-2, n = 10), OVX treated with LY294002 (OVX + LY294002, n = 10), and OVX treated with GLP-2 plus LY294002 (OVX + GLP-2 + LY294002, n = 10). A IHC staining of FGF23 in the distal femur and corresponding MOD analysis. B Relative mRNA expression of FGF23 in femurs. C Serum FGF23 levels measured by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: FGF23 levels in cell culture supernatants were measured using a mouse FGF23 ELISA Kit (CSB-EL008629MO, Cusabio) following the manufacturer’s protocol.

Techniques: Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: GLP-2 alleviates postmenopausal osteoporosis by acting on osteoblasts through the PI3K/AKT/FOXO1 signalling pathway to downregulate FGF23 expression

doi: 10.1186/s13018-026-06661-2

Figure Lengend Snippet: GLP-2 downregulates FGF23 expression by activating GLP-2R/PI3K/AKT signalling in osteoblasts. A Western blot analysis of AKT and p-AKT levels in osteoblasts with or without shRNA-GLP-2R interference. B Relative mRNA expression of FGF23 in osteoblasts treated with or without shRNA-GLP-2R. C Western blot analysis of AKT and p-AKT levels in cells treated with or without LY294002. D Relative mRNA expression of FGF23 following LY294002 intervention. E Western blot analysis of AKT and p-AKT levels in cells treated with or without MK-2206. F Relative mRNA expression of FGF23 after MK-2206 treatment. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: FGF23 levels in cell culture supernatants were measured using a mouse FGF23 ELISA Kit (CSB-EL008629MO, Cusabio) following the manufacturer’s protocol.

Techniques: Expressing, Western Blot, shRNA

Journal: Journal of Orthopaedic Surgery and Research

Article Title: GLP-2 alleviates postmenopausal osteoporosis by acting on osteoblasts through the PI3K/AKT/FOXO1 signalling pathway to downregulate FGF23 expression

doi: 10.1186/s13018-026-06661-2

Figure Lengend Snippet: GLP-2 regulates FGF23 expression via the inactivation of FOXO1. A Western blot analysis of FOXO1 and p-FOXO1 levels in osteoblasts treated with or without GLP-2. B Immunofluorescence staining of FOXO1 in osteoblasts (green), counterstained with DAPI (blue). C Relative mRNA expression of FGF23 in osteoblasts treated with the FOXO1 inhibitor AS1842856. D Verification of FOXO1 overexpression by Western blot after PCDH-FOXO1 transfection. E Effect of FOXO1 overexpression on FGF23 mRNA levels in osteoblasts with or without GLP-2 treatment. F Effect of FOXO1 overexpression on the FGF23 concentration in the cell culture supernatant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: FGF23 levels in cell culture supernatants were measured using a mouse FGF23 ELISA Kit (CSB-EL008629MO, Cusabio) following the manufacturer’s protocol.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Over Expression, Transfection, Concentration Assay, Cell Culture

Journal: Journal of Orthopaedic Surgery and Research

Article Title: GLP-2 alleviates postmenopausal osteoporosis by acting on osteoblasts through the PI3K/AKT/FOXO1 signalling pathway to downregulate FGF23 expression

doi: 10.1186/s13018-026-06661-2

Figure Lengend Snippet: Mechanistic diagram illustrating how GLP-2 downregulates FGF23 to ameliorate postmenopausal osteoporosis via the GLP-2R/PI3K/AKT/FOXO1 signalling pathway

Article Snippet: FGF23 levels in cell culture supernatants were measured using a mouse FGF23 ELISA Kit (CSB-EL008629MO, Cusabio) following the manufacturer’s protocol.

Techniques:

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: FGF19 is associated with CRC liver metastasis (CRCLM) and poor prognosis in CRC patients. A) Seventeen cell subsets were identified by analysis of the scRNA‐Seq data from 6 patients with CRCLM. B) Analysis of the cellular communication in tumor samples from patients with CRCLM. C) The Venn diagram illustrating the overlap of interacting genes between CRC cells and CAFs, as well as the highly expressed genes in CRC from the TCGA database. D) Volcano plot showing the differential expression of genes, and nine of the interacting genes exhibited a marked increase in expression. E) Correlation analysis between FGF19 expression and CRC patient survival (n = 597). F) The FGF19 expression in colon adenocarcinoma (COAD) patients from the TCGA database (Normal, n = 41; Primary tumor, n = 286). G) Representative images of the luminance signals from the CRC (nonmetastasis), CRCLM, and CRC pulmanary metastasis (CRCPM) animal models. CT26‐luciference‐expressing (CT26‐luc) cells were injected into the caecum, spleen, and tail vein of mice to establish the CRC (CRC‐no metastasis), CRCLM, and CRCPM animal models, respectively. The luminance signals were detected by using the IVIS Lumina XRMS Series III instrument at the end of the experiment. H) Representative images and the luminance signals of the colon, lung, and liver tissues of the mice in CTL (n = 7), CRC (CRC‐no metastasis) (n = 6), CRCPM (n = 7), and CRCLM (n = 7) animal models. I) The levels of FGF15 in the serum of CRC, CRCLM, or CRCPM model mice were detected using the ELISA assay (CTL, n = 7; CRC, n = 6; CRCPM, n = 7; CRCLM, n = 7). J) IF staining showing the expression of FGF19 in the clinical samples using the microarray assay (n = 42). Data are shown as Mean ± SD. For I, ** p < 0.01, versus CTL.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Quantitative Proteomics, Expressing, Injection, Enzyme-linked Immunosorbent Assay, Staining, Microarray

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: CRC cells release FGF19 (FGF15) which activates HSCs and promotes HSCs‐to‐CAFs differentiation. A) The experimental setup for the control (upper panel) and experimental (lower panel) conditions for the conditioned medium (CM) systems. B) Experimental setup for the control (upper panel) and experimental (lower panel) coculture systems. C,D) The protein levels of α‐SMA and FAP in the CM system (C) and SW620‐LX‐2 coculture system (D) were determined by the Western blotting; and the quantitative results are shown in the right panel. E,F) The protein levels of α‐SMA and FAP in the CM system (E) and CT26‐JS1 coculture system (F) were determined by the Western blotting; and the quantitative results are shown in the right panel. G) Effects of the recombinant FGF19 protein on the protein levels of FAP and α‐SMA in LX‐2 cells. H) Effects of FGF15 recombinant protein on the protein levels of FAP and α‐SMA in JS1 cells. I,J) Effects of infigratinib, a FGFR inhibitor on the protein levels of FAP and α‐SMA in SW620 CM system (I) or CT26 CM system (J). Data are shown as Mean ± SD, n = 3. For C, E: * p < 0.05, ** p < 0.01 versus HSCs CM. For D, F‐H: * p < 0.05, ** p < 0.01 versus the corresponding CTL. For I‐J: * p < 0.05, ** p < 0.01 versus HSCs CM; # p < 0.05, ## p < 0.01 versus CRC CM.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Control, Western Blot, Recombinant

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: FGF‐activated HSCs increase CRC cells migration. A,B) Representative images of SW620 (A) and CT26 (B) cells migration in the CRC cells‐HSCs coculture system (left panel); and the quantitative data were analyzed using Image J software (right panel). Photographs were taken 24 h after treatment. C) Representative images of cell migration in the SW620, SW620‐LX‐2 coculture, and FGF19‐treated coculture systems (left panel); and the quantitative data were analyzed using Image J software (right panel). D) Representative images of cell migration in the CT26, CT26‐JS1 coculture, or FGF15 treated coculture system (left panel); and quantitative results were analyzed using Image J software (right panel). E) Representative images of cell migration in SW620, SW620‐LX‐2 coculture, and infigratinib‐treated coculture system (left panel); and quantitative results were analyzed using Image J software (right panel). F) Representative images of cell migration in the CT26, CT26‐JS1 coculture, and infigratinib‐treated coculture systems (left panel). All the quantitative results were analyzed using Image J software (right panel). Data are shown as Mean ± SD from three independent experiments, n = 3. For A‐D: * p < 0.05, ** p < 0.01 versus the corresponding CTL. For E‐F: * p < 0.05, ** p < 0.01 versus the corresponding CTL; # p < 0.05, ## p < 0.01 versus CRC cells‐HSCs coculture group.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Migration, Software

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: Effects of FGF15 and ANGPTL4 in mouse models of different degrees of CRCLM. A) Expression levels of MMP2 and MMP9 in the liver tissues of each group were detected by IHC staining. B,C) Protein levels of FGF15 in the liver tissues (B) and tumor sites (C) were determined by Western blotting. D) The serum level of FGF15 was determined by the ELISA assay. E) Expression of FGFR4, a FGF15 receptor in the liver tissues of each group was detected by IHC staining. F) Expression levels of α‐SMA and FAP in the liver tissues of each group were detected by IHC staining. G,H) Protein levels of α‐SMA in the liver tissues with tumors were determined by using Western blotting (G), and quantitative results were analyzed using Image J software (H). I) mRNA levels of FAP in the liver tissues with tumors were determined using RT‐qPCR analysis. J) Protein levels of ANGPTL4 in the liver tissues were determined by Western blotting (left panel), and the quantitative data were analyzed using Image J software (right panel). K,L) The liver tissue homogenates (K) and the serum (L) levels of ANGPTL4 were determined by the ELISA assay. M) Localization of FGF15 in CTCs within the liver of CRCLM mice by using the mIF analysis. N) Localization of ANGPTL4 in the liver tissue CAFs in CRCLM mouse model determined using the mIF analysis. Data are shown as Mean ± SD, n = 6. For B, D, H‐L, * p < 0.05, ** p < 0.01 versus CTL. For C, ** p < 0.01 versus 5.0 × 10 5 group.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Expressing, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay, Software, Quantitative RT-PCR

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: The effects of FGF15 and ANGPTL4 in CRCLM mouse models with different time points. A) Timeline for the establishment of the mouse CRCLM model. CT26‐luc cells were injected into the spleens of BALB/c mice, then the mice were sacrificed at different time points. B) Representative images of live tumor‐bearing mice with tumors. C) Representative fluorescence signal imaging of the liver tissues. D,E) Quantitative results of the mice fluorescence intensity (D) and the liver fluorescence intensity (E) were analyzed using the Living Image software 4.4. F–H) Representative IHC staining of FGF15 and FGFR4 (F) MMP2 and MMP9 (G), α‐SMA, and FAP (H) in the liver tissues of mice in each group. I) Localization of ANGPTL4 in CAFs of liver tissues in CRCLM mouse model by using the mIF analysis. J) Protein levels of ANGPTL4 in the liver tissues with tumors were determined by using Western blotting (upper panel); and quantitative results were quantified and analyzed using Image J software (lower panel). K,L) Liver tissue homogenates (K) and the serum (L) levels of ANGPTL4 were determined by the ELISA assay. Data are shown as Mean ± SD. n = 6. * p < 0.05, ** p < 0.01 versus CTL.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Injection, Fluorescence, Imaging, Software, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: FGF19‐Activated Hepatic Stellate Cells Release ANGPTL4 that Promotes Colorectal Cancer Liver Metastasis

doi: 10.1002/advs.202413525

Figure Lengend Snippet: The FGF15/ANGPTL4 axis is involved in the progression of CRCLM. A,B) Representative images of IHC staining for FGF15 and FGFR4 (A), and for α‐SMA and FAP (B) in the liver tissues of mice in each group. C) The protein levels of ANGPTL4 in tumor‐bearing liver tissues were determined by using Western blotting (upper panel); and quantitative results were analyzed using Image J software (lower panel). D) Representative live‐animal imaging images of mice with tumors. CT26‐luc‐shNC cells and CT26‐luc‐shFGF15 cells were inoculated into the hepatic portal vein of BALB/c mice, respectively. Mice were randomly divided into 3 groups, including the sham, shNC, and shFGF15 groups. E) Quantitative results of the fluorescence intensity were analyzed using Living Image software 4.4. F) Representative H&E‐stained images of liver tissues in each group of mice. G) Representative images of IHC for MMP2 and MMP9 in mouse liver tissues from each group. H) Diagram showing that the FGF19/ANGPTL4 axis mediates the interaction between CRC cells and HSCs, and promotes CRC liver metastasis. Data are shown as Mean ± SD, n = 6. ** p < 0.01 versus Sham; # p < 0.05, ## p < 0.01 versus shNC.

Article Snippet: Infigratinib, human angiopoietin‐related 4 protein (ANGPTL4, HEK293, His), and mouse FGF15 Protein (His‐SUMO) were purchased from MedChemExpress (MCE, NJ, USA).

Techniques: Immunohistochemistry, Western Blot, Software, Imaging, Fluorescence, Staining