Journal: Biochemical and biophysical research communications

Article Title: Overexpression of Snail in retinal pigment epithelial triggered epithelial-mesenchymal transition.

doi: 10.1016/j.bbrc.2014.02.119

Figure Lengend Snippet: Fig. 3. Overexpression of Snail in ARPE-19 cells induced EMT. ARPE-19 cells were transfected with pReceiver-Snail or pReceiver-control for 48 h. QRT-PCR and Immunoblotting were used to examine the expression of Snail, E-cadherin, ZO-1, a-SMA and fibronectin. (A) QRT-PCR analysis showed the increased Snail, fibronectin and a- SMA mRNA expression and decreased E-cadherin and ZO-1 mRNA expression. ⁄⁄P < 0.01 vs pReceiver-control. (B) Immunoblotting confirmed the expression of these EMT markers at protein levels.

Article Snippet: The primary antibodies used were as follows: 1:500 E-cadherin antibody and 1:1000 fibronectin antibody (R&D systems, Inc., USA), 1:1000 Snail antibody (Abcam Ltd., Cambridge, USA), 1:1000 a-SMA antibody (Sigma–Aldrich, MO, USA), 1:1000 ZO-1 antibody (Invitrogen, Carlsbad, CA), and 1:5000 GAPDH (Good HERE, Hangzhou, China).

Techniques: Over Expression, Transfection, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Transcriptome analysis revealed the central role of the ECM-PI3K pathway in pancreatic cancer metastasis. (A) Volcano plot of differentially expressed genes (DEGs), showing 248 DEGs between pancreatic cancer tissues (PANC) and adjacent normal tissues (CTR), with 153 genes upregulated and 95 genes downregulated. (B) Hierarchical clustering heatmap of the top 100 DEGs, demonstrating differences in gene expression patterns between PANC and CTR tissues. (C) Results of KEGG functional enrichment analysis, indicating that the DEGs are primarily involved in signaling pathways such as ECM-receptor interaction, Cytoskeleton in muscle cells, Focal adhesion, and P13K/AKT signaling pathway. (D) GO analysis results, showing significant enrichment of DEGs in processes related to adhesion and ECM. (E, F) CytoHubba analysis results, revealing FN1 as a core hub gene in the P13K/AKT signaling pathway.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Gene Expression, Functional Assay, Protein-Protein interactions

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Single-cell sequencing identifies specific expression patterns of PI3K-related genes including FN1 in fibroblasts of metastatic samples. (A) UMAP plot displaying the sub-cell types in the primary pancreatic cancer (stiu) and liver metastasis (meta) samples from the single-cell RNA sequencing dataset GSE156405 . (B) UMAP plot showing the expression patterns of different groups in primary pancreatic cancer (stiu) and liver metastasis (meta) samples. (C) Cell proportion plot illustrating the distribution of different cell types in stiu and meta samples. (D) Bubble plot representing the relationship between different sub-cell types and characteristic gene expression. The color of the bubbles ranges from white to blue, representing gene expression percentages of 0%, 25%, 50%, and 75%, respectively. The size of the bubbles indicates the average expression level, ranging from a minimum to a maximum representing average expression values from 0 to 2. (E) KEGG enrichment analysis of DEGs in alveolar epithelial cells. (F) KEGG enrichment analysis of DEGs in fibroblasts. (G) Violin plot showing the expression patterns of PI3K-related genes (FN1, THBS2, COL1A1, COL1A2, and COL6A3) in different cell types and groups in fibroblasts.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Sequencing, Expressing, RNA Sequencing, Gene Expression

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: Core mechanisms of CAFs in pancreatic cancer metastasis. (A) UMAP plot displaying fibroblast subpopulations by cluster. (B) UMAP plot showing the distribution of fibroblast subpopulations by group. (C) Proportion plot illustrating the cell proportions by cell subpopulation and group. (D) Identification of the CAFs subpopulation among fibroblast subpopulations. (E) UMAP visualization analysis, displaying the expression patterns of FN1, THBS2, COL1A1, COL1A2, and COL6A3 in the stiu and meta groups.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: CAF-derived FN1 promotes invasion and migration of pancreatic cancer cells. (A) Schematic diagram of the CAFs-PANC1 co-culture model. (B) ELISA results showing changes in IL-6, IL-8, and MMP2 levels secreted by CAFs before and after TGF-β induction. (C) ELISA results demonstrating changes in FN1 levels secreted by CAFs before and after treatment with FN1 neutralizing antibody (FN1-Ab). (D) Transwell migration assay showing changes in the number of migrated cells in the FN1-Ab group compared to the control group. (E) Statistics of migration rates from the Transwell migration assay. (F) Transwell invasion assay showing changes in the number of invading cells in the FN1-Ab group compared to the control group. (G) Statistics of invasion rates from the Transwell invasion assay.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Derivative Assay, Migration, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Transwell Migration Assay, Control, Transwell Invasion Assay

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: The FN1-ITG-PI3K/AKT axis promotes invasion and migration of pancreatic cancer cells. (A, B) GEPIA database analysis revealing a significant positive correlation between FN1 expression and the expression of key genes in the PI3K/AKT pathway (PIK3CA, AKT1). (C, D) Western blot results showing changes in phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab group. The Western blot results demonstrated the changes in the phosphorylation levels of p-AKT and p-PI3K in the FN1-Ab groups of PANC-1 and BXPC-3 cells. (E, F) Statistical data corresponding to (C, D) . (G, H) mRNA expression changes of integrin genes (ITGA2, ITGB4, ITGA3) before and after treatment with an integrin inhibitor (ITG-Inh). The changes in mRNA expression of integrin genes ITGA2 , ITGB4 , and ITGA3 in PANC-1 and BXPC-3 cells before and after treatment with the integrin inhibitor ITG-Inh. (I, J) Western blot results demonstrating changes in protein levels of p-AKT and p-PI3K in the ITG-Inh group compared to the control group (ITG-Con). The Western blot results indicated that in PANC-1 and BXPC-3 cells, compared with the control group (ITG-CON), the protein levels of p-AKT and p-PI3K in the ITG-Inh group were altered. (K, L) Statistical data corresponding to (I, J) . (M, N) Changes in invasion ability after combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh). The changes in invasive capacity following the combined inhibition of PI3K and integrins (PI3K-Inh + ITG-Inh) in PANC-1 and BXPC-3 cells. (O, P) Statistical data corresponding to (M, N) .

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Migration, Expressing, Western Blot, Phospho-proteomics, Control, Inhibition

Journal: Frontiers in Oncology

Article Title: FN1 from cancer-associated fibroblasts orchestrates pancreatic cancer metastasis via integrin-PI3K/AKT signaling

doi: 10.3389/fonc.2025.1595523

Figure Lengend Snippet: High FN1 expression is associated with poor prognosis and an immunosuppressive microenvironment. (A) Clinical data analysis showing the relationship between FN1 expression and patient survival. (B) TIMER2.0 displaying the correlation between FN1 expression and M2 macrophage infiltration in pancreatic cancer (PAAD). (C) TIMER2.0 showing the correlation between FN1 expression and Treg cell infiltration in PAAD. (D) Mechanism diagram illustrating how FN1 activates the PI3K/AKT pathway by binding to integrin receptors, thereby promoting the invasion and metastasis of pancreatic cancer cells.

Article Snippet: FN1 neutralizing antibody (FN1-Ab) was purchased from R&D Systems (Cat# MAB19182).

Techniques: Expressing, Binding Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Kefir peptides mitigate bleomycin-induced pulmonary fibrosis in mice through modulating oxidative stress, inflammation and gut microbiota.

doi: 10.1016/j.biopha.2024.116431

Figure Lengend Snippet: Fig. 5. Pretreatment KPs attenuates the expression of fibrotic-related genes in the bleomycin-induced pulmonary fibrosis mouse model. Mice pre-treated with water or KPs for 4 days following intratracheal injection of bleomycin for 21 days. Lung tissues were harvested to detect fibrotic-related mRNA, including (A) Fn1, (B) Col3a1, (C) Col1a1, (D) Timp1, and (E) Ctgf expression by quantitative RT-PCR (n=5 mice per group). Values are normalized to the β-actin gene and are expressed relative to the control group. (F) Western blot analysis was performed to analyze the expression of Collagen I and fibronectin in the lung tissues from each group (n=4–5 mice per group). *p < 0.05, **p < 0.01, and ***p < 0.001. All the statistical information and details of tests for each figure are available in the supple mentary files.

Article Snippet: After transferring to PVDF membranes, the membranes were blocked with 5% nonfat dry milk in TBS-T (0.1% Tween-20 in TBS buffer) for 1 h. Proteins were then probed overnight at 4 ◦C with the following antibodies at appropriate dilutions: 1:2000 dilution of Fibronectin and Collagen type I (Proteintech) and 1:25,000 dilution of β-actin (Novus Biologicals, Centennial, CO, USA).

Techniques: Expressing, Injection, Quantitative RT-PCR, Control, Western Blot

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: Knockout of GSDMD alleviated renal tubule morphological changes in OXO/adenine-induced mice and attenuated fibrosis (A and B) Histological changes in renal tubules after HE (A) and Masson (B) staining in different groups. Scale bar 50 μm and 100μm. (C and D) Western blot showing α-SMA levels in renal tissue from different groups (n = 6 animals per group). (C and E) Western blot showing E-cadherin levels in renal tissue from different groups (n = 5 animals per group). (F and G) RT‒PCR analysis of fibronectin (F) and collagen I (G) levels in renal tissue from different groups (n = 5 animals per group). (H) RT‒PCR analysis of IL-1β levels in renal tissue from different groups (n = 6 animals per group). Data for (D)-(H) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (D–H). WT, wild type. Norm-diet, normal diet. UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Knock-Out, Staining, Western Blot

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: The primer sequences used in RT‒PCR

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques:

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: GSDMD encouraged IL-1β expression and fibrosis in NRK-52E cells in vitro (A–D) HK-2 cells were stimulated with uric acid for 48 h. Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) levels in HK-2 cells in the different groups (n = 3 per group). (E–H) NRK-52E cells were stimulated with uric acid for 48 h. Western blot showing cleaved-caspase-1 (E and F), caspase-1 (E and G) and RIG-I (E and H) levels in NRK-52E cells in the different group (n = 6 per group). (I–P) NRK-52E cells were transfected with GSDMD-siRNA or NC-siRNA and treated with UA for 48 h. Western blot showing GSDMD (I and J) and GSDMD-N (I and K) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). (L) IL-1β level in culture medium (n = 6 per group). (M–P) Western blot showing α-SMA (M and N), E-cadherin (M and O) and fibronectin (M and P) levels after UA treatment with or without GSDMD knockdown (n = 6 per group). Data for (B–D), (F–H), (J–L) and (N–P) are presented as mean ± SEM. Student’s t test for (B–D) and (F–H). One-way ANOVA with Student-Newman-Keuls test for (J–L) and (N–P). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Expressing, In Vitro, Western Blot, Transfection, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet: RIG-I-siRNA decreased HUA-induced fibrosis and pyroptosis molecules expressions in NRK-52E cells in vitro NRK-52E cells were transfected with RIG-I-siRNA or NC-siRNA and treated with UA for 48 h. (A–D) Western blot showing caspase-1 (A and B), GSDMD (A and C) and RIG-I (A and D) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (E–G) Western blot showing cleaved-caspase-1 (E and F) and GSDMD-N (E and G) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). (H) IL-1β level in culture medium (n = 6 per group). (I–K) Western blot showing fibronectin (I and J) and α-SMA (I and K) expressions after UA treatment with or without RIG-I knockdown (n = 6 per group). Data for (B–D), (F–H) and (J–K) are presented as mean ± SEM. One-way ANOVA with Student-Newman-Keuls test for (B–D), (F–H) and (J–K). UA, uric acid.

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: In Vitro, Transfection, Western Blot, Knockdown

Journal: iScience

Article Title: Gasdermin D promotes hyperuricemia-induced renal tubular injury through RIG-I/caspase-1 pathway

doi: 10.1016/j.isci.2023.108463

Figure Lengend Snippet:

Article Snippet: Anti-Fibronectin antibody , Cell Signaling Technology , Cat# 63779.

Techniques: Recombinant, H&E Stain, Staining, Activity Assay, Transfection, Magnetic Beads, Enzyme-linked Immunosorbent Assay, Lysis, Bradford Protein Assay, Bicinchoninic Acid Protein Assay, Negative Control, Software, Fluorescence, Microscopy

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 2. Effects of A-485 on the profibrotic phenotype induced by TGF-β1 in HK-2 cells. (A) The representative western blot images and quantification of FN, Col-I, E- cad and α-SMA in cultured HK-2 cells (n = 3). (B) Micrographs of α-SMA and E-cad immunofluorescence staining in HK-2 cells (n = 6). (C-D) The quantification of α-SMA and E-cad immunofluorescence staining in HK-2 cells. Values are expressed as means ± SEM. *p < 0.05, * *p < 0.01. FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Cell Culture, Immunofluorescence, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A-485 alleviates fibrosis and apoptosis in kidney by disrupting tandem activation of acetylation and phosphorylation on STAT3.

doi: 10.1016/j.biopha.2025.118217

Figure Lengend Snippet: Fig. 4. Effects of A-485 on UUO-induced profibrotic proteins in mouse kidney. (A-B) The representative western blot images and quantification of FN, Col-I, E-cad and α-SMA in mice (n = 6). (C) Representative micrographs of FN, Col-I and E-cad immunohistochemical staining in mice. (D-F) The quantification of FN, Col-I and E- cad expression detected by immunohistochemical staining (n = 5). Values are expressed as means ± SEM. * *p < 0.01. sham, sham operation; UUO: unilateral ureteral obstruction; FN, fibronectin; Col-I, collagen I; E-cad, E-Cadherin; α-SMA, α-smooth muscle actin.

Article Snippet: The antibodies used in the present study are listed as below: antibodies against Bcl-2 (ab196495), Collagen I (ab34710); antibodies against STAT3 (10253–2-AP), Bax (50599–2-Ig), α-SMA (14395–1-AP), Fibronectin (15613–1-AP), E-Cadherin (20874–1-AP) and GAPDH (10494–1-AP) (Proteintech, Chicago, IL, USA); antibodies against Acetyl-STAT3 (Lys685) (#2523), Cleaved Caspase-3 (#9661), p300 (#67621), CBP(#74384SF), Acetylated-Lysine (#9441), PhosphoSTAT3 (S727) (#49081) and Phospho-STAT3 (Y705) (#9145) (Cell Signaling Technology, Beverly, MA, USA); antibodies against NDUFS1 (ER1803–08) and SDHB (ER1803–63) (Hua’an Biotechnology, Hangzhou, China); antibody against COX4 (sc-517553) (Santa Cruz Biotechnology, Santa Cruz, CA); goat anti-rabbit or mouse IgG-HRP secondary antibodies, TRITC-labelled goat anti-rabbit secondary antibody and FITC-labelled goat anti-rabbit or mouse IgG secondary antibodies (Zhongshan Goldenbridge Biotechnology, Beijing, China); Inhibitor A-485 (1889279–16–6) (GlpBio, Montclair, USA).

Techniques: Western Blot, Immunohistochemical staining, Staining, Expressing

Journal: Cancer cell

Article Title: Cancer metastasis is accelerated through immunosuppression during Snail-induced EMT of cancer cells.

doi: 10.1016/j.ccr.2009.01.023

Figure Lengend Snippet: Figure 1. snail-Transduced Murine Mela- noma B16-F10 Cells with EMT-like Charac- teristics (A) Left: decrease of epithelial markers and increase of mesenchymal markers in Snail trans- fectants detected by RT-PCR. Right: the intensity of snail, E-cadherin, and fibronectin was normal- ized to GAPDH expression as a control. (B) Decreased E-cadherin and increased fibro- nectin in the Snail transfectants with spindle and dendritic shapes (immunocytochemistry; scale bars = 0.5 mm). (C) Decreased proliferation and adhesion and increased invasion of Snail transfectants (n = 3, mean ± SD). (D) Suppressed growth of Snail transfectants in vivo. C57BL/6 mice were implanted subcutane- ously (s.c.) with tumor cells (n = 5, mean ± SD). (E) Enhanced in vivo metastatic ability of Snail transfectants. Mice were implanted intravenously (i.v.) with tumor cells, and the numbers of nodules in lungs of the mice were counted on day 14 after tumor implantation (n = 5, mean ± SD). (F) Decreased survival of mice with Snail transfec- tants. Mice were implanted i.v. with tumor cells. Data shown in each panel are representative of four independent experiments.

Article Snippet: After treatment with Cytofix/Cytoperm solution (BD Pharmingen), tumor cells were stained with anti-Snail mAb (Santa Cruz Biotechnology), anti-E-cadherin mAb (BD Pharmingen), anti-fibronectin mAb (R&D Systems), or the appropriate isotype control antibodies.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Immunocytochemistry, In Vivo, Tumor Implantation

Journal: Cancer cell

Article Title: Cancer metastasis is accelerated through immunosuppression during Snail-induced EMT of cancer cells.

doi: 10.1016/j.ccr.2009.01.023

Figure Lengend Snippet: Figure 7. Immunosuppression Induced by snail-Transduced HumanMelanomaHS294TCells with EMT-likeCharacteristics (A) Left: decrease of the epithelial marker E-cadherin and increase of the mesenchymal marker fibronectin and immunosuppressive cytokines (TGF-b, IL10, and TSP1) in the Snail transfectants detected by RT-PCR. Right: the intensity of snail, E-cadherin, and fibronectin was normalized to GAPDH expression as a control. (B) Decrease of E-cadherin and increase of fibronectin in the Snail transfectants with spindle and dendritic shapes (immunocytochem- istry; scale bars = 0.5 mm). (C) EMT-like functional changes of Snail transfectants: decreased proliferation, decreased adhesion, and increased invasion (n = 3, mean ± SD). (D) Increase of CD4+Foxp3+ cells and decrease of CD11c+HLA-DR+

Article Snippet: After treatment with Cytofix/Cytoperm solution (BD Pharmingen), tumor cells were stained with anti-Snail mAb (Santa Cruz Biotechnology), anti-E-cadherin mAb (BD Pharmingen), anti-fibronectin mAb (R&D Systems), or the appropriate isotype control antibodies.

Techniques: Marker, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Functional Assay