Journal: Frontiers in Immunology

Article Title: Mode of Proximal Tubule Damage: Differential Cause for the Release of TFF3?

doi: 10.3389/fimmu.2016.00122

Figure Lengend Snippet: Densitometric analysis of proteome array reactivities for TFF3 (A) and characteristic markers of hypoxia (FABP1, VEGF) and inflammation (CXCL16) (B) using cell lysates from HK-2 cells exposed to hypoxia/nutrient starvation (HNS) and replenishment (HNSR) for 24 and 48 h . Densities obtained are shown in (C) . These were normalized to a reference (pos) and the background was subtracted. Data are presented as mean ± SD and were compared using Student’s multiple t -test. p < 0.05 was considered statistically significant. GraphPad Prism 6.07 was used for statistical analysis (GraphPad Software, La Jolla, CA, USA). HIF-1α abundance is increased at the early replenishment time point when normalized to a housekeeping protein β-actin and compared to the normoxic control (D) . Normalized abundance of TFF3 (7 kDa) in cell lysates from HK-2 cells exposed to hypoxia/nutrient starvation (HNS) and replenishment for 24 and 48 h (E) . NDL, normalized density to loading control (Western blot).

Article Snippet: TFF3 antibody was purchased from Biorbyt Ltd. (Cambridge, UK) and used at 1:500 with DAKOmate Envision detection kit (1:40).

Techniques: Software, Control, Western Blot

Journal: Frontiers in Immunology

Article Title: Mode of Proximal Tubule Damage: Differential Cause for the Release of TFF3?

doi: 10.3389/fimmu.2016.00122

Figure Lengend Snippet: The human kidney biomarker array (R&D Systems) was used to analyze TFF3 abundance in lysates from cells exposed to immunoglobulin λ light chain (LC) and fatty acid free human serum albumin (FAF-HSA) for 72 h in comparison with control cells (A) . Densities were normalized to a reference (pos), background subtracted and are expressed as ±SD. (B) Abundance of TFF3 (in relation to β-actin) in cell lysates from HK-2 cells exposed 5 mg/ml immunoglobulin λ light chain (LC) or fatty acid-free human serum albumin (FAF-HSA) for 72 h (Western blot). (C) ELISA of TFF3 in the supernatants of HK-2 stimulated with 5 mg/ml LC and FAF-HSA. Untreated cells were used as control. Two independent experiments for the distinct stimulations for 24 and 72 h are shown.

Article Snippet: TFF3 antibody was purchased from Biorbyt Ltd. (Cambridge, UK) and used at 1:500 with DAKOmate Envision detection kit (1:40).

Techniques: Biomarker Discovery, Comparison, Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Comparison of TFF1 and TFF3 mRNA in normal and cancer tissue.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: mRNA expression of TFF1 and TFF3 in cancer and normal tissues and the association with survival. (A) mRNA expression of TFF1 or TFF3 in cancer tissues was higher than that in normal tissues (P=0.034 or P=0.007). (B) Survival analysis revealed that a higher expression of TFF3 was associated with a worse overall survival, while TFF1 expression demonstrated no association with patient survival (P<0.001 and P=0.125). * P<0.05 and ** P<0.01 vs. normal group. TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Correlation between mRNA expression of TFF1 and TFF3 in patients with CRC.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Association between TFF1 or TFF3 mRNA expression and patient clinicopathological factors.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Expressing, Cell Differentiation

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Significant predictive factors (univariate and multivariate) for overall survival in Cox proportional-hazard analysis.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques:

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Promoter methylation sites and levels of the TFF1 and TFF3 genes in cancer and normal tissues. (A) The length of promoter region of the TFF1 gene for detection was 489 bp, 13 units of CpG sites could be found and 10 units were measured successfully. (B) The length of 372 bp was detected for the TFF3 gene and 7 out of 8 units were analyzed favorably. The different colors represent the relative methylation changes in 10% increments (yellow, 0%; blue, 100% methylated). TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Methylation

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Comparison of average methylation level of TFF1 and TFF3 between normal and cancer groups.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Methylation

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Comparison of average methylation levels in different CPG sites of TFF3 between normal and cancer groups.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Methylation

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Correlation between the methylation level and mRNA expression of TFF3.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Methylation, Expressing

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Correlation analysis between the promoter methylation level and mRNA expression of TFF1 or TFF3. No apparent variation trends were observed from the scatter diagrams for TFF1 or TFF3. TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Methylation, Expressing

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: The overexpression of TFF3 promotes the proliferation, migration and invasion capacities of the HT29 cells. (A) Both western blot analysis and RT-qPCR revealed that TFF3 expression in the HT29 and LoVo cells were the lowest and highest, respetively. (B) TFF3 expression was found to be enhanced significantly at the mRNA and protein level following transfection with the TFF3 expression plasmid. (C) CCK-8 assay revealed that the overexpression of TFF3 markedly increased the proliferation of the HT29 cells. (D) Transwell chamber assay displayed that the migration and invasion of the HT29 cells was markedly enhanced following the overexpression of TFF3. (E) The migration of the HT29 cells was also markedly increased according to wound healing assay. (F) Compared with the control group, the pIRES2-TFF3-transfected cells exhibited less pseudopodia (A), a reduced number of villi (B) and fewer intercellular junctions (C and D). An increased number of polygonal or spindle like shaped cells in the TFF3 overexpression group was also observed (B). * P<0.05 and ** P <0.01 vs. control (HIEC) group or the empty vector (Vector). TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Over Expression, Migration, Western Blot, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, CCK-8 Assay, Transwell Chamber Assay, Wound Healing Assay

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: Knockdown of TFF3 notably reduces the proliferation, migration and invasion of LoVo cells. (A) TFF3 expression was suppressed by shRNA transfection, as shown by both western blot analysis and RT-qPCR. (B) CCK-8 assay revealed that the silencing of TFF3 significantly decreased the proliferation of the LoVo cells. (C) The migration and invasion of Lovo cells were markedly suppressed following the silencing of TFF3, as shown by Transwell chamber assay. (D) Scratch wound healing assay demonstrated that the migration of the LoVo cells was markedly reduced by the silencing of TFF3. * P<0.05 and ** P<0.01 vs. shvector. TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Migration, Expressing, shRNA, Transfection, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Transwell Chamber Assay, Wound Healing Assay

Journal: International Journal of Oncology

Article Title: TFF3 and TFF1 expression levels are elevated in colorectal cancer and promote the malignant behavior of colon cancer by activating the EMT process

doi: 10.3892/ijo.2019.4854

Figure Lengend Snippet: TFF3 induces the EMT process in colorectal cancer cells. (A) Western blot analysis demonstrated that the enforced expression of TFF3 in HT29 cells increased the protein expression of Twist1, Snail and Vimentin, while it suppressed E-cadherin expression. (B) The downregulation of TFF3 expression in the Lovo cells decreased the protein expression of Twist1, Snail and Vimentin, while it enhanced E cadherin expression. (C) The same results were observed at the mRNA level when TFF3 was overexpressed or silenced. (D) EMT-like morphological changes in cells following the overexpression or silencing of TFF3. The morphology of the HT29 cells changed from a round or volmer-weber mode into a triangular or spindle form following the overexpression of TFF3. LoVo cells displayed a reversed morphological change from the scattered cord-like form to the round, rectangular or clustered pattern after the silencing of TFF3. #P<0.05 and ## P<0.01vs. empty vector (Vector) or shVector. TFF, trefoil factor.

Article Snippet: Subsequently, the membranes were probed with primary antibodies against TFF1 (1:500, ab92377) TFF3 (1:300, ab108599) and Twist1 (1:500, ab50581), E cadherin (1:500, ab40772), Vimentin (1:500, ab92547), Snail (1:500, ab53519), β-actin (1:2,500, ab8226) (all from Abcam) in blocking solution overnight at 4°C.

Techniques: Western Blot, Expressing, Over Expression, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: p53, miR-34a and EMP1—Newly Identified Targets of TFF3 Signaling in Y79 Retinoblastoma Cells

doi: 10.3390/ijms20174129

Figure Lengend Snippet: TFF3 overexpression enhances p53 transcriptional activity and protein expression and increases miR-34a expression with downstream reduction of EMP1 in Y79 RB cells. ( A ) Quantitative Real-time PCR confirmation of TFF3 lentiviral overexpression (Trefoil factor family peptide 3 (TFF3)) in Y79 cells compared to control cells (ctr). ( B ) Luciferase assays were performed with Y79 cells transiently transfected with TFF3 or empty vector control (ctr) in addition to wild-type PG13-Luc (wt PG13) or mutant MG15-Luc (mut MG15). Forced TFF3 expression leads to an increased luciferase signal upon p53 promotor activation in Y79 cells. ( C ) Western blot analysis showing increased p53 and TFF3 protein levels after TFF3 overexpression (TFF3). The indicated intensity ratios of p53 and TFF3 protein levels relative to β-actin levels were calculated using ImageJ software. ( D , E ) Quantitative real-time PCR analysis of miR-34a and EMP1 expression levels in Y79 cells compared to control cells after lentiviral TFF3 overexpression (ctr). Values are means of at least 3 independent experiments ± SEM. * p -value < 0.05, ** p -value < 0.01; statistical differences compared to the control group calculated by Student’s t -test or one-way ANOVA and Newman-Keuls post test.

Article Snippet: Primary antibodies used (incubated overnight at 4 °C): EMP1 (1:1000): ab230445, Abcam; p53 (1:400): sc-71817, Santa Cruz; TFF3 (1:500): TA307376, Origene; ß-actin (1:1000): #4967, Cell Signalling.

Techniques: Over Expression, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Control, Luciferase, Transfection, Plasmid Preparation, Mutagenesis, Activation Assay, Western Blot, Software

Journal: Rhinology journal

Article Title: Expression of Trefoil factor family peptides in the nasal allergic mucosa

doi: 10.4193/rhino11.221

Figure Lengend Snippet: Figure 1. Immunostaining for TFF1and TFF3 in the nasal mucosa. (A) TFF1: The staining was positive in epithelial goblet cells and ciliated cells, and

Article Snippet: Sections were then labeled with primary antibodies to TFF1 (rabbit, 1:100 dilution, Novocastra Laboratories Ltd.) and TFF3 (rabbit, 1:50 dilution, Protein Tech Group, Inc.) overnight at MUC5AC in the submucosal glands (D) (original magnification 200×).

Techniques: Immunostaining, Staining

Journal: Rhinology journal

Article Title: Expression of Trefoil factor family peptides in the nasal allergic mucosa

doi: 10.4193/rhino11.221

Figure Lengend Snippet: Figure 2. Double-staining for MUC5AC (red), TFF1 (green), and TFF3 (green) in the nasal mucosa. TFF1 co-localized with MUC5AC in the epithelium (A),

Article Snippet: Sections were then labeled with primary antibodies to TFF1 (rabbit, 1:100 dilution, Novocastra Laboratories Ltd.) and TFF3 (rabbit, 1:50 dilution, Protein Tech Group, Inc.) overnight at MUC5AC in the submucosal glands (D) (original magnification 200×).

Techniques: Double Staining

Journal: Rhinology journal

Article Title: Expression of Trefoil factor family peptides in the nasal allergic mucosa

doi: 10.4193/rhino11.221

Figure Lengend Snippet: Figure 3. Double-staining for MUC5B (red), TFF1 (green), and TFF3 (green) in the nasal mucosa. TFF1 co-localized with MUC5B in the epithelium (A)

Article Snippet: Sections were then labeled with primary antibodies to TFF1 (rabbit, 1:100 dilution, Novocastra Laboratories Ltd.) and TFF3 (rabbit, 1:50 dilution, Protein Tech Group, Inc.) overnight at MUC5AC in the submucosal glands (D) (original magnification 200×).

Techniques: Double Staining

Journal: Rhinology journal

Article Title: Expression of Trefoil factor family peptides in the nasal allergic mucosa

doi: 10.4193/rhino11.221

Figure Lengend Snippet: Figure 6. Correlation of mRNA expression values for TFF3 and MUC5B in the nasal mucosa of allergic (AR+) and nonallergic (AR-) patients.

Article Snippet: Sections were then labeled with primary antibodies to TFF1 (rabbit, 1:100 dilution, Novocastra Laboratories Ltd.) and TFF3 (rabbit, 1:50 dilution, Protein Tech Group, Inc.) overnight at MUC5AC in the submucosal glands (D) (original magnification 200×).

Techniques: Expressing

Journal: British Journal of Nutrition

Article Title: Phytic acid improves intestinal mucosal barrier damage and reduces serum levels of proinflammatory cytokines in a 1,2-dimethylhydrazine-induced rat colorectal cancer model

doi: 10.1017/s0007114518001290

Figure Lengend Snippet: Fig. 4. Real-time PCR analysis of mucin 2 (MUC2) (a) and trefoil factor 3 (TFF3) (b) mRNA expression levels. The normal colon tissues were selected in the control group (CG) and the colon tumour tissues were selected in the other groups. β-Actin was used as a loading control. MG, model group; PA, phytic acid; LPG, low-PA-dose group; MPG, middle-PA-dose group; HPG, high-PA-dose group. **P<0·01, CG compared with the MG; †† P< 0·01, PA treatment groups compared with the MG.

Article Snippet: The following antibodies were used in the Western blot analysis: rabbit anti-MUC2 (Abcam), claudin-1 (Novus), TFF3 (Novus) and E-cadherin (Danvers).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Control

Journal: British Journal of Nutrition

Article Title: Phytic acid improves intestinal mucosal barrier damage and reduces serum levels of proinflammatory cytokines in a 1,2-dimethylhydrazine-induced rat colorectal cancer model

doi: 10.1017/s0007114518001290

Figure Lengend Snippet: Fig. 6. Western blot analysis of mucin 2 (MUC2) and trefoil factor 3 (TFF3) protein expression. The normal colon tissues were selected in the control group (CG) and the colon tumour tissues were selected in the other groups. β-Actin was used as a loading control. (a) Representative blots from one of three separate experiments; (b) relative band intensities of MUC2 based on densitometry; (c) relative band intensities of TFF3 based on densitometry. MG, model group; PA, phytic acid; LPG, low-PA-dose group; MPG, middle-PA- dose group; HPG, high-PA-dose group. ** P < 0·01, CG compared with the MG; †† P < 0·01, PA treatment groups compared with the MG.

Article Snippet: The following antibodies were used in the Western blot analysis: rabbit anti-MUC2 (Abcam), claudin-1 (Novus), TFF3 (Novus) and E-cadherin (Danvers).

Techniques: Western Blot, Expressing, Control