Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects of low-serum adaptation on growth and protein expression in stably lactoferrin-overexpressing mammary alveolar cells

doi: 10.3389/fbioe.2026.1731548

Figure Lengend Snippet: Immunofluorescence images of MAC-T cells. (A) Nucleus staining of MAC-T cells; (B) Cytokeratin 18 (CK18) staining; (C) Combined nucleus and CK18 staining. KSR: Knockout™ serum replacement. ITS: insulin–transferrin–selenium. Scale bar = 200 μm.

Article Snippet: After washed with pre-cooled PBS for 5 min, cells were fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.1% Triton-X-100 for 1 h, and blocked with 5% BSA at room temperature for 2 h. Then primary antibody CK18 (Bioss, bs2043R) diluted at 1:100 was added, followed by overnight incubation at 4 °C and incubation with a fluorescent secondary antibody (Proteintech, RGAR004) at 1:500 dilution in the dark for 2 h. The cells were washed thrice with PBS and stained with DAPI (Beyotime, C1106) ( ).

Techniques: Immunofluorescence, Staining, Knock-Out

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

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Expression of 3 hub genes (A) Gene expression of KCNK1, PCDH1 and SDR16C5. (B) Expression of KCNK1 in a single cell. (C) Expression of PCDH1 in a single cell. (D) Expression of SDR16C5 in a single cell.

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Expressing, Gene Expression

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Related assays on the expression of hub genes. (A–C) mRNA expression of KCNK1, PCDH1 and SDR16C5 in HPNE and BXPC3. (D) mRNA expression of PCDH1 in HPNE and multiple PDAC cell lines. (E) Matched pairs of PDAC tissues and adjacent non-neoplastic pancreatic tissues were stained with PCDH1-specific antibodies for immunohistochemical analysis. Representative images of tissues and tumor-infiltrating cells are shown (100×, scale bar = 100 μm; 400×, scale bar = 20 μm). The MOD of PCDH1 is obtained by analyzing the photo-optical density with IPP software. (F) ROC curve of PCDH1 in PDAC. (G) Protein-protein interaction network of PCDH1. (ns: P >0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Expressing, Staining, Immunohistochemical staining, Software

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Correlation between PCDH1 and immune infiltration. (A) Correlation between PCDH1 and immune score. (B) Correlation between PCDH1 and stromal score. (C) Correlation between PCDH1 and ESTIMATE score. (D) Correlation between PCDH1 and immune cells. (* P < 0.05, ** P < 0.01).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques:

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Chemosensitivity of PCDH1. (A) Correlation between GDSC drug sensitivity and PCDH1 mRNA expression (B) Correlation between CTRP drug sensitivity and PCDH1 mRNA expression.

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Knockdown of PCDH1 inhibited the proliferation of PDAC. (A) Efficiency of knocking down mRNA of PCDH1 si-RNA in BXPC3. (B) Efficiency of knocking down mRNA of PCDH1 si-RNA in SW1990. (C, E) MTT assays reveal the proliferation ability of control group and PCDH1 knockdown group in cell line BXPC3 and SW1990. (D, F) Colony formation assays reveal the proliferation ability of control group and PCDH1 knockdown group in cell line BXPC3 and SW1990. (ns: P >0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Knockdown, Control

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Knockdown of PCDH1 inhibited the migration of PDAC. (A) Transwell assays show the migration ability of the control group and the PCDH1 knockdown group in the BXPC3 cell line. (Scale bar = 50μm) (B) Transwell assays show the migration ability of the control group and the PCDH1 knockdown group in the SW1990 cell line. (Scale bar = 50μm) (C) Wound healing assays exhibit the migration ability of the control group and the PCDH1 knockdown group in the BXPC3 cell line. (Scale bar = 100μm) (D) Wound healing assays exhibit the migration ability of the control group and the PCDH1 knockdown group in the SW1990 cell line. (Scale bar = 100μm) (ns: P >0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Knockdown, Migration, Control

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: Knockdown of PCDH1 inhibited the tumor stemness of PDAC cell lines. (A) Relative expression of CD24 mRNA in the control group and the PCDH1 knockdown group. (B) Relative expression of CD133 mRNA in the control group and the PCDH1 knockdown group. (C) Detection of CD24 + cells in the control and PCDH1 knockdown groups via flow cytometry in BXPC3 and SW1990 cell lines. (D) Detection of CD133 + cells in the control and PCDH1 knockdown groups via flow cytometry in BXPC3 and SW1990 cell lines. (E, F) Percentage of CD24 + cells between the control group and the PCDH1 knockdown group (G, H) Percentage of CD133 + cells between the control group and the PCDH1 knockdown group. (I, J) Sphere formation assay was performed to detect sphere formation ability of the control group and the PCDH1 knockdown group in BXPC3 and SW1990. (K, L) Diameter of tumor spheres at 24 h, 48 h and 5 days in the control group and the PCDH1 knockdown group.(Scale bar = 500μm) (ns: P >0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Knockdown, Expressing, Control, Flow Cytometry, Tube Formation Assay

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: PCDH1 promotes proliferation and migration of PDAC through activation of PI3K/Akt pathway. (A) Heatmap of differential expression genes between the NC and si-PCDH1 groups. (B) KEGG enrichment analysis demonstrates pathways associated with differential expression genes. (C, D) Expression levels of mRNA for six molecules in the PI3K/Akt pathway were detected by qRT-PCR. (*** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Migration, Activation Assay, Quantitative Proteomics, Expressing, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: PCDH1 promotes proliferation and migration of PDAC through activation of PI3K/Akt pathway. (A, B) MTT assay was used to detect the proliferation ability of the control, IGF-1, si-PCDH1, and si-PCDH1 + IGF-1 groups. (C–E) A colony formation assay was performed to detect the proliferation ability of the four groups. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Migration, Activation Assay, MTT Assay, Control, Colony Assay

Journal: Frontiers in Oncology

Article Title: PCDH1 facilitates migration, proliferation, and stemness of pancreatic cancer cells through PI3K-Akt signaling

doi: 10.3389/fonc.2025.1696695

Figure Lengend Snippet: PCDH1 promotes proliferation and migration of PDAC through activation of PI3K/Akt pathway. (A) Transwell assays present the migration capacity of the control, IGF-1, si-PCDH1, and si-PCDH1 + IGF-1 groups. (Scale bar = 100μm) (B, C) Counts of cell migration in four groups. (D, F) Wound healing assays exhibit the migration ability of the four groups. (Scale bar = 100μm) (E, G) Rate of cell migration in four groups. (** P < 0.01, *** P < 0.001, **** P < 0.00011).

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-PCDH1 primary antibody (bs-111111R, Bioss Biotechnology, China) at the manufacturer-recommended dilution.

Techniques: Migration, Activation Assay, Control