Journal: Cells

Article Title: Dibutyltin(IV) and Tributyltin(IV) Derivatives of meso -Tetra(4-sulfonatophenyl)porphine Inhibit the Growth and the Migration of Human Melanoma Cells

doi: 10.3390/cells8121547

Figure Lengend Snippet: Kinetics of the integrins and FAK expression in A375 cells treated with (Bu 2 Sn) 2 TPPS and (Bu 3 Sn) 4 TPPS. The kinetics of ( A ) integrin β1, ( B ) integrin β3 adhesion receptors and of ( C ) FAK and pFAK expression was analysed in A375 melanoma cells treated with 250 nM of (Bu 2 Sn) 2 TPPS (A, B and C, left and right panels) and with 80 nM of (Bu 3 Sn) 4 TPPS ( A , B and C , middle and right panels) for 24 h, 48 h and 72 h, through western blot experiments. The analysis of the β-actin expression ( A , B and C , lower panels) was used to confirm the equal protein loading. The error bars indicate the standard deviation. The Student’s t -test was used to the analysis of statistical significance: * p < 0.05 was considered significant; ** p < 0.01 highly significant; *** p < 0.001 very highly significant ( A , B and C , right panels).

Article Snippet: The anti-pTyr-397 FAK motif, integrin β1, ICAM-1 and MCAM mouse monoclonal antibodies were obtained from BD Biosciences (Lexington, KY, USA).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Cells

Article Title: Dibutyltin(IV) and Tributyltin(IV) Derivatives of meso -Tetra(4-sulfonatophenyl)porphine Inhibit the Growth and the Migration of Human Melanoma Cells

doi: 10.3390/cells8121547

Figure Lengend Snippet: Kinetics of the integrins and FAK expression in HT-144 cells treated with (Bu 2 Sn) 2 TPPS and (Bu 3 Sn) 4 TPPS. The kinetics of ( A ) integrin β1, ( B ) integrin β3 adhesion receptors and of ( C ) FAK and pFAK expression was analysed in HT-144 melanoma cells treated with 200 nM of (Bu 2 Sn) 2 TPPS ( A , B and C , left and right panels) and with 60 nM of (Bu 3 Sn) 4 TPPS ( A , B and C , middle and right panels) for 24 h, 48 h and 72 h, through western blot experiments. The analysis of the β-actin expression ( A , B and C , lower panels) was used to confirm the equal protein loading. The error bars indicate the standard deviation. The Student’s t -test was used to the analysis of statistical significance: * p < 0.05 was considered significant; ** p < 0.01 highly significant; *** p < 0.001 very highly significant ( A , B and C , right panels).

Article Snippet: The anti-pTyr-397 FAK motif, integrin β1, ICAM-1 and MCAM mouse monoclonal antibodies were obtained from BD Biosciences (Lexington, KY, USA).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: Enhanced cytotoxicity against cholangiocarcinoma by fifth-generation chimeric antigen receptor T cells targeting integrin αvβ6 and secreting anti-PD-L1 scFv

doi: 10.1186/s12967-025-06453-y

Figure Lengend Snippet: A20 CAR5 T cells infiltrate integrin αvβ6 + /PD-L1 + KKU-213A spheroids. A Representative images were captured to visualize the infiltration of A20 CAR T cells into the spheroids. In brief, 4 × 10 3 cells of KKU-213A labeled with CellTracker ™ Blue 4-chloromethyl-6,8-difluoro-7-hydroxycoumarin (CMF 2 HC) were co-cultured with either NT T, A20 CAR4 T or A20 CAR5 T cells labeled with CFSE (Thermo Fisher Scientific) at an E:T ratio of 2:1. B Histograms represent the averaged intensity profiles of blue CMF 2 HC KKU-213A spheroids and green CFSE T cells signals in z-stack cross-sections of spheroids defied by ImageJ. C T cell infiltration was quantified by calculating the area under the curve (AUC) of the green CFSE signals. Scale bar = 200 μm. Statistical significance was determined using one-way ANOVA followed by Tukey’s post hoc test (** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: The cells were then incubated overnight at 4 °C with mouse anti-integrin β6 antibody (1:100 dilution, Clone 442.5C4; Merck Millipore, Massachusetts, USA) and rabbit anti-PD-L1 antibody (1:100 dilution, Clone 28–8, Abcam, Cambridge, UK).

Techniques: Labeling, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Enhanced cytotoxicity against cholangiocarcinoma by fifth-generation chimeric antigen receptor T cells targeting integrin αvβ6 and secreting anti-PD-L1 scFv

doi: 10.1186/s12967-025-06453-y

Figure Lengend Snippet: Expression of integrin αvβ6 and PD-L1 in cholangiocarcinoma (CCA) cell lines. A Immunofluorescence analysis showing the expression of integrin αvβ6 (green) and PD-L1 (red) in three CCA cell lines: KKU-055, KKU-100, and KKU-213A. The scale bar represents 50 μm. B Immunoblot analysis detecting PD-L1 ( ~ 55 kDa) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 37 kDa) as a loading control (upper panel). Densitometry quantification of PD-L1 relative to GAPDH was performed using ImageJ software (lower panel). C Flow cytometry histograms displaying the surface expression of integrin αvβ6 and ( E ) PD-L1, with isotype controls shown in light gray. Quantification of ( D ) integrin αvβ6- and ( F ) PD-L1-positive cells is presented as percentages. Data are shown as mean ± standard error of the mean (SEM) from three independent experiments (N = 3). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test (* p < 0.05, *** p < 0.001, **** p < 0.0001)

Article Snippet: The cells were then incubated overnight at 4 °C with mouse anti-integrin β6 antibody (1:100 dilution, Clone 442.5C4; Merck Millipore, Massachusetts, USA) and rabbit anti-PD-L1 antibody (1:100 dilution, Clone 28–8, Abcam, Cambridge, UK).

Techniques: Expressing, Immunofluorescence, Western Blot, Control, Software, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: Enhanced cytotoxicity against cholangiocarcinoma by fifth-generation chimeric antigen receptor T cells targeting integrin αvβ6 and secreting anti-PD-L1 scFv

doi: 10.1186/s12967-025-06453-y

Figure Lengend Snippet: Schematic illustrating the mechanism of A20 CAR5 T cells, designed to mimic the natural activation process of T cells. The integrin αvβ6-targeted CAR recognizes and binds to integrin αvβ6 present on cancer cells, while PD-L1, an immune checkpoint molecule, inhibits T cell activity by interacting with PD-1 expressed on the T cell surface. Full T cell activation occurs when integrin αvβ6 is recognized, and PD-L1 is neutralized by the secreted anti-PD-L1 scFv. Without this neutralization, PD-L1 remains active, preventing full T cell activation and reducing the immune response

Article Snippet: The cells were then incubated overnight at 4 °C with mouse anti-integrin β6 antibody (1:100 dilution, Clone 442.5C4; Merck Millipore, Massachusetts, USA) and rabbit anti-PD-L1 antibody (1:100 dilution, Clone 28–8, Abcam, Cambridge, UK).

Techniques: Activation Assay, Activity Assay, Neutralization

Journal: Arthritis Research & Therapy

Article Title: Prostaglandin E2 synthesis in cartilage explants under compression: mPGES-1 is a mechanosensitive gene

doi: 10.1186/ar2024

Figure Lengend Snippet: Over-release of prostaglandin E2 (PGE 2 ) in compressed costal cartilage explants is the result of mechanical stress. (a) Implication of the mechanoreceptor integrin α5β1 in PGE 2 over-release in compressed cartilage explants. Mouse costal cartilage explants treated with either the β1 non-blocking antibody VMA1997 or the α5β1 blocking antibody AB1950 at 2.5 μg/ml were compressed (C) or not compressed (NC) for 4 h. Results are normalized to the mean not-compressed control (cont) value. Data are the mean ± SEM of 2 independent experiments with n = 2/group/experiments, analyzed in duplicate. ***p < 0.001 versus control NC, *p < 0.05 versus control C. (b) Increased PGE 2 release in compressed costal cartilage explants is not due to the cytokine IL-1. Mouse costal cartilage explants treated with the IL-1 receptor antagonist (IL1-Ra) at 100 ng/ml were compressed (C) or not compressed (NC) for 4 h. Results are normalized to the mean not compressed control value. Data are the mean ± SEM of 2 independent experiment with n = 2/group/experiments, analyzed in duplicate. *p < 0.05 versus control NC.

Article Snippet: Anti-goat fibronectin receptor (integrin α5β1) blocking polyclonal antibody (AB1950) was purchased from Euromedex for Chemicon Inc. (Strasbourg, France) and rat anti-mouse β1 subunit of VLA1 integrins non-blocking monoclonal antibody (VMA 1997), was purchased from AbCys SA for Chemicon Inc. (Souffelweyersheim, France).

Techniques: Blocking Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

doi: 10.1186/s13046-018-0783-6

Figure Lengend Snippet: SNCG protein is associated with β1 integrin and activates β1 integrin. a - b . Coimmunoprecipitation. Cell membrane proteins of HCT116 cells were collected and subjected to immunoprecipitated (IP) using anti-SNCG ( a ), anti-SNCG or anti-β1 integrin antibody ( b ). The IP proteins or total cell lysates were analyzed by Western blot. Normal IgG served as the negative control. c . Far-Western blot analysis. HCT116 cells were transfected with control siRNA (lane 1-2), and specific siRNA-β1-2 (lanes 3-4) for 48 h. Cells were treated without (lane 1, 3) or with 1 μmol/L rhSNCG (lane 2, 4). Cell lysates were subjected to SDS-PAGE and transferred to NC membrane. β1 integrin (prey protein) on the membrane is detected with SNCG (bait protein). More SNCG was associated with membrane β1 integrin in SNCG-treated cells than that in the control cells (lane 1, 2). Correspondingly, less SNCG were detected in β1 integrin knock-down cells than that in control cells (lane 2, 4). d - e , Effect of concentration and time treatment of SNCG on activated β1 integrin. HCT116 cells were stimulated with GST or GST-SNCG at various concentrations for 60 min ( d ) or at fixed concentration (1 μmol/L) for various times ( e ). Cell lysates were analyzed with the HUTS-21 mAb recognizing the activated form of β1 integrin. f , GST-SNCG treatment (1 μmol/L) upregulated activated β1 integrin subunit in HCT116 and SW480 cells. g , Colocalization of SNCG with F-actin. HCT116 cells grown on coverslips were transiently transfected with control siRNA or β1-specific siRNA-2. After 72 h, cells were treated with GST or GST-SNCG (1 μmol/L) for 60 min. Cells were fixed and stained with anti-SNCG (red) and FITC-Phalloidin (green). Colocalization of SNCG and F-actin was shown in yellow. Nuclei were counterstained with DAPI (blue). Scale bars, 5 μm

Article Snippet: Mouse anti-human β1 integrin monoclonal antibody (mAb) (specifically recognizing the active conformation) is from BD Biosciences.

Techniques: Immunoprecipitation, Western Blot, Negative Control, Far Western Blot, Transfection, SDS Page, Concentration Assay, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

doi: 10.1186/s13046-018-0783-6

Figure Lengend Snippet: Integrin β1 is required for enhancement of SNCG on tumor cell migration and invasion. a - b , The functional blocking antibody for β1 integrin subunit (5, 10, 20 μg/mL) was added in the upper compartment of migration or invasion chambers stimulated with or without GST-SNCG (1 μmol/L) for 24 h for migration ( a ) or 48 h for invasion ( b ). c - d , HCT116 cells were transfected with control siRNA, and β1-specific siRNA-2 for 48 h. then cells were treated with or without GST-SNCG (1 μmol/L) for 24 h for migration ( c ) or 48 h for invasion ( d ). e - f , HCT116 cells were treated with PBS or 200 μmol/L RGD for 30 min and then treated with or without GST-SNCG (1 μmol/L) for 24 h for migration ( e ) or 48 h for invasion ( f ). Graphed data represent the mean ± SE from at least six 200-power field for each condition, two-sample t-test. g , HCT116 cells were transfected with control siRNA, and β1-specific siRNA-2 for 48 h. Then cells were treated with or without GST-SNCG (1 μmol/L) for 30 min and cell lysates were analyzed by Western blot. h , HCT116 cells were treated with PBS or 200 μmol/L RGD for 30 min. Then cells were treated with or without GST-SNCG (1 μmol/L) for 30 min and cell lysates were analyzed by Western blot

Article Snippet: Mouse anti-human β1 integrin monoclonal antibody (mAb) (specifically recognizing the active conformation) is from BD Biosciences.

Techniques: Migration, Functional Assay, Blocking Assay, Transfection, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

doi: 10.1186/s13046-018-0783-6

Figure Lengend Snippet: FAK is essential for SNCG-enhanced tumor cell migration and invasion. a - b , HCT116 cells were transfected with control siRNA and FAK-specific siRNA-2, -3 for 48 h. Then cells were treated with or without GST-SNCG (1 μmol/L) for migration ( a ) or invasion ( b ). c - d , HCT116 cells were treated with PBS, 50 μmol/L FAK inhibitor 14 for 30 min and then treated with or without GST-SNCG (1 μmol/L) for migration ( c ) or invasion ( d ). Graphed data represent the mean ± SE from at least six 200-power field for each condition, two-sample t-test. e - f , HCT116 cells were transfected with control siRNA (lane 1-2), and FAK-specific siRNA-2 (lane 3-4) and -3 (lane 5-6) for 72 h. cells were treated with or without GST-SNCG (1 μmol/L) for 30 min and cell lysates were analyzed for activated and total β1 integrin ( e ) or activated and total FAK ( f )

Article Snippet: Mouse anti-human β1 integrin monoclonal antibody (mAb) (specifically recognizing the active conformation) is from BD Biosciences.

Techniques: Migration, Transfection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

doi: 10.1186/s13046-018-0783-6

Figure Lengend Snippet: SNCG is an indicator of adverse prognosis and positively correlates with activated β1 integrin, p-FAK (Y 397 ) in CRC tissues. a - b , Kaplan-Meier estimation of disease-free survival (DFS) for stage I-II ( a ) and III-IV ( b ) colorectal adenocarcinoma patients according to SNCG levels. c , Correlations of SNCG levels in CRC tissues with post-operative recurrence and status. d , Representative blots from three independent experiments were presented. Protein levels of SNCG, activated β1 integrin, and p-FAK (Y 397 ) in clinical colon cancer tissue samples were evaluated by Western blot analysis. In order to increase the reproducibility, HCT116 cell lysates were used in each blot as the internal control ( c ) to minimize the effect of band intensity variation. GAPDH was used as the loading control. e - g , Correlation between the relative protein levels of activated β1 integrin levels and p-FAK (Y 397 ) ( e ), SNCG and active β1 integrin ( f ), and SNCG and p-FAK (Y 397 ) ( g ) were plotted as a scatter plots

Article Snippet: Mouse anti-human β1 integrin monoclonal antibody (mAb) (specifically recognizing the active conformation) is from BD Biosciences.

Techniques: Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Extracellular gamma-synuclein promotes tumor cell motility by activating β1 integrin-focal adhesion kinase signaling pathway and increasing matrix metalloproteinase-24, -2 protein secretion

doi: 10.1186/s13046-018-0783-6

Figure Lengend Snippet: Exogenously added SNCG remodels the microenvironment of tumor cells and increases MMP-2 activity by β1 integrin. a , Antibody array screening of CM from GST and GST-SNCG-treated HCT116 cells. b , CM (left panel) and whole cell lysates (right panel) from HCT116 and SW480 cells treated with or without GST-SNCG (1 μmol/L) were subjected to Western blot analysis. Representative blots from three independent experiments were presented. c - d , HCT116 cells were treated with diluent, 50 μmol/L MMP-2 inhibitor for 40 min, then 1 μmol/L GST or GST-SNCG was added in the cell medium for migration ( c ) or invasion ( d ) assay. Migrated or invaded cells were quantitated after 24 h or 48 h, respectively. Error bars, SE of three determinations. e - f , Western blot and gelatin zymography analysis. CM from HCT116 cells treated with GST or GST-SNCG (1 μmol/L) in β1 integrin knock-down ( e ) or RGD-treated cells ( f ) was analyzed by gelatin zymography with FBS as the positive control, and secreted protein levels were analyzed by Western blot

Article Snippet: Mouse anti-human β1 integrin monoclonal antibody (mAb) (specifically recognizing the active conformation) is from BD Biosciences.

Techniques: Activity Assay, Ab Array, Western Blot, Migration, Zymography, Positive Control

Journal: JCI Insight

Article Title: Targetable purinergic receptors P2Y 12 and A2b antagonistically regulate bladder function

doi: 10.1172/jci.insight.122112

Figure Lengend Snippet: Deletion of P2Y12 and A2b result in enlarged (A) and reduced (B) bladder size, respectively. (C–E) H&E staining images from bladder sections of wild-type, P2Y12-KO, and A2b-KO mice, respectively. (F–H) Immunostaining of Ki67 from bladder sections of wild-type (n = 5), P2Y12-KO (n = 5), and A2b-KO (n = 5) mice, respectively. Positive nuclear staining per bladder section is quantitated in I. (J–L) Integrin β1 immunostaining of bladder sections of wild-type (n = 186 cells), P2Y12-KO (n = 225 cells), and A2b-KO (n = 185 cells) mice, respectively. (M–O) Enlarged images of the white boxes seen in J, K, and L, respectively. (P) Schematic diagram indicating where the bladder was sectioned for staining, and how only bladder smooth muscle (BSM) cells containing nuclei were measured for their cross-section area, which serves as an index of BSM cell size. (Q) BSM cell cross-sectional area was quantitated for each model. (R and S) Quantitative RT-PCR data for c-fos and c-jun mRNA expression in mouse bladder from wild-type (n = 3), P2Y12-KO (n = 3), and A2b-KO (n = 3) mice. (T) Western blot images of c-fos and c-jun protein expression in mouse bladder from wild-type (n = 6), P2Y12-KO (n = 6), and A2b-KO (n = 6) mice, which are quantitated by densitometry and normalized to β-actin in U and V. Data are shown as box and whiskers, whiskers are from minimum to maximum, Student’s t test is used to compare between wild-type and KO animals; *P < 0.05.

Article Snippet: Fixed tissue was cryoprotected, frozen, sectioned, and incubated with rabbit polyclonal anti-Ki67 antibody (catalog ab15580, abcam) and purified rat anti-mouse β1 integrin antibody (catalog 550531, BD Bioscience) (1:100 dilution) overnight at 4°C.

Techniques: Staining, Immunostaining, Quantitative RT-PCR, Expressing, Western Blot