Journal: Microbiology Spectrum

Article Title: Lactococcus petauri LZys1 modulates gut microbiota, diminishes ileal FXR-FGF15 signaling, and regulates hepatic function

doi: 10.1128/spectrum.01716-24

Figure Lengend Snippet: L. petauri LZys1 decreases ileal-derived circulating FGF15 and downregulates hepatic FGFR4-FXR-mediated BA metabolism and fatty acid oxidation. ( A ) The expression levels of genes associated with the FXR/FGF15 pathway in the ileum, normalized to β-actin ( n = 5, per group). ( B ) Serum FGF15 concentration ( n = 4, per group). ( C ) Expression of genes associated with the FXR/FGF15 pathway in the liver, normalized to β-actin ( n = 6, per group). ( D ) Representative Western blot images illustrating the level of FGFR4 and FXR protein in the liver ( n = 6, per group). ( E ) The relative grayscale values of Western blot images in ( D ). ( F ) The expression of genes associated with BA secretion and synthesis of the conjugated BAs in the liver, normalized to β-actin ( n = 6, per group). ( G ) The expression of genes associated with oxidation of fatty acids in the liver, normalized to β-actin ( n = 6, per group). ( H ) The expression of profibrotic growth factors and proinflammatory cytokine genes in the liver, normalized to β-actin ( n = 6, per group). Data ( A–H ) are expressed as mean ± SEM and compared by two-sample t -test. * P < 0.05, ** P < 0.01.

Article Snippet: The membranes were blocked with 5% defatted milk in Tris-buffered saline with Tween 20 (TBST) at room temperature for 2 h and then incubated with FXR (Santa Cruz Biotechnology, sc-25309, 1:100), FGFR4 (Santa Cruz Biotechnology, sc-136988, 1:100), and β-actin (CST, 12262, 1:100).

Techniques: Derivative Assay, Expressing, Concentration Assay, Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

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

doi: 10.1002/advs.202413525

Figure Lengend Snippet: Figure 5. 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 × 105 group.

Article Snippet: Chemicals and Reagents: Antibodies against GAPDH (Cat# sc-32233), α-SMA (Cat# sc-53142), FGF19 (Cat# sc-390621), FGF15 (Cat# sc514647), MMP9 (Cat# sc-13520), FGFR4 (Cat# sc-136988), and ANGPTL4 (Cat# sc-373761) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against MMP2 (Cat# 87809) and goat antirabbit HRP (Cat# 7074) were purchased from Cell Signaling Technology (CST, MA, USA); antibody against EpCAM (Cat# bs-1513R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (Bioss, Beijing, China); antibody against FAP (Cat# A11572) was purchased from ABclonal Technology Co., Ltd. (ABclonal, Wuhan, China); goat anti-mouse IgG HRP (Mu Biotech, Cat# 125035) was purchased from Mu Biotechnology, Inc. (Mu Biotech, Guangzhou, China).

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

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

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

doi: 10.1002/advs.202413525

Figure Lengend Snippet: Figure 6. 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 deter- mined 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: Chemicals and Reagents: Antibodies against GAPDH (Cat# sc-32233), α-SMA (Cat# sc-53142), FGF19 (Cat# sc-390621), FGF15 (Cat# sc514647), MMP9 (Cat# sc-13520), FGFR4 (Cat# sc-136988), and ANGPTL4 (Cat# sc-373761) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against MMP2 (Cat# 87809) and goat antirabbit HRP (Cat# 7074) were purchased from Cell Signaling Technology (CST, MA, USA); antibody against EpCAM (Cat# bs-1513R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (Bioss, Beijing, China); antibody against FAP (Cat# A11572) was purchased from ABclonal Technology Co., Ltd. (ABclonal, Wuhan, China); goat anti-mouse IgG HRP (Mu Biotech, Cat# 125035) was purchased from Mu Biotechnology, Inc. (Mu Biotech, Guangzhou, China).

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

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

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

doi: 10.1002/advs.202413525

Figure Lengend Snippet: Figure 7. 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: Chemicals and Reagents: Antibodies against GAPDH (Cat# sc-32233), α-SMA (Cat# sc-53142), FGF19 (Cat# sc-390621), FGF15 (Cat# sc514647), MMP9 (Cat# sc-13520), FGFR4 (Cat# sc-136988), and ANGPTL4 (Cat# sc-373761) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against MMP2 (Cat# 87809) and goat antirabbit HRP (Cat# 7074) were purchased from Cell Signaling Technology (CST, MA, USA); antibody against EpCAM (Cat# bs-1513R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (Bioss, Beijing, China); antibody against FAP (Cat# A11572) was purchased from ABclonal Technology Co., Ltd. (ABclonal, Wuhan, China); goat anti-mouse IgG HRP (Mu Biotech, Cat# 125035) was purchased from Mu Biotechnology, Inc. (Mu Biotech, Guangzhou, China).

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