bioss  (Bioss)

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: SNS activated FXR to modulate LDs transport. A – C Protein expression levels of GPAT4 and LDAH in liver tissues. D IHC staining demonstrates GPAT4 expression levels and localization in liver tissues. E , F ORO staining demonstrated successful model establishment, and SNS and OCA reduced LDs deposition in cells. G – L Western blot analysis was performed to confirm that SNS and OCA activated FXR and inhibited GPAT4 expression in vitro. Scale bars, 50 μm, 100 μm and 200 μm. Data are presented as mean ± SD (n = 5). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, In Vitro

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: Knockdown of GPAT4 reduced LDs deposition, while knockdown of FXR increased GPAT4 expression. A , B The knockdown efficiency of GPAT4 was detected by Western blot assay. C The knockdown efficiency of GPAT4 was verified by PCR experiments, and 1505 was selected as the effective GPAT4 knockdown for subsequent experiments. D , E ORO staining was used to analyze the effect of GPAT4 knockdown on LDs deposition in cells. F , G , H Western blot and PCR experiments were used to screen and verify the knockdown efficiency of FXR, and 786 was selected for subsequent experiments. I , J , K After knocking down FXR, western blot and PCR experiments were performed to detect the expression level of GPAT4. L , M ORO staining was used to analyze the effect of FXR knockdown on LDs deposition in cells. Scale bar, 50 μm. Data are expressed as mean ± SD (n = 3). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Knockdown, Expressing, Western Blot, Staining

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: Database predictions of FXR binding to GPAT4, validated by DL. A , B The UCSC database was used to predict correlation scores and their corresponding p-values. C Predicted binding sequence of FXR. D The JASPAR database analyzed the binding correlation and potential binding sites between FXR and GPAT4. E Schematic diagram of constructing the plasmid for overexpressing FXR. F Schematic diagram of FXR binding sites in the three predicted GPAT4 promoter regions. G , H Western blot was used to detect the expression level of FXR in cells transfected with the plasmid overexpressing FXR. I DL was used to detect the binding of transcription factor FXR to the GPAT4 promoter region. Data are presented as mean ± SD (n = 3). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Binding Assay, Sequencing, Plasmid Preparation, Western Blot, Expressing, Transfection

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: The active components of SNS could bind to FXR and GPAT4. A – G Visualization of the binding of Glycyrrhizin, Hesperidin, Liquiritin, Neohesperidin, Isorhamnetin, Peoniflorin and Nobiletinto to FXR. H – N Visualization of the binding of Glycyrrhizin, Hesperidin, Liquiritin, Neohesperidin, Isorhamnetin, Peoniflorin and Nobiletinto to GPAT4. O – Q The RMSD, Rg and RMSF values of the FXR-Liquiritin complex over time. R – T The RMSD, Rg and RMSF values of the GPAT4-Neohesperidin complex over time

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Binding Assay

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: As illustrated in the figure, SNS activates hepatic FXR, inhibits the LDs transport protein GPAT4, and modulates the expression of proteins involved in lipolysis and lipophagy, including P62, Beclin1, LC3Ⅰ/Ⅱ, HSL, ATGL, and MAGL. Activation of hepatic FXR inhibits GPAT4 expression. Conversely, when FXR is inhibited, LDs deposition in liver cells worsens, leading to feedback upregulation of GPAT4 expression. The FXR-GPAT4 axis was validated using a DL

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Expressing, Activation Assay