β1 integrin antibody  (Developmental Studies Hybridoma Bank)

Journal: International Journal of Cancer

Article Title: Arp2/3 complex and β1 integrin drive an invasive front through extracellular matrix adaptation in pancreatic cancer

doi: 10.1002/ijc.70376

Figure Lengend Snippet: β1 integrin signaling facilitates Arp2/3‐dependent migration on type I collagen. (A) Volcano plot of RNAseq results comparing Arpc4 KO versus control in 8025 and R254 cells. Functional annotation of downregulated genes (8025 cells, n = 379; R254 cells, n = 328) after Arpc4 knock‐out in murine PDAC cells. Top 10 annotations are shown. Reference genome: GRCm39. (B) Western blot shows expression levels of Arpc4, β1 integrin and p‐MLC2 in control and Arpc4 KO cells, Hsp90 as a loading control. (C) Panels depict single‐cell trajectories on plates coated with type I collagen with treatment of 100 μg/mL IgG or β1‐Integrin antibody. Quantitative analysis for migrated distance and velocity of cells in each group. (D) Representative phase‐contrast pictures show the morphology of control 8025 cells in type I collagen (left, scale bars: 200 μm) with treatment of 100 μg/mL IgG or β1‐Integrin antibody at various time points. Quantitative analysis of size for the structures formed by PDAC cells (right). p ‐values for β1‐Integrin antibody vs. IgG are 0.0002 (Day 3), <0.0001 (Day 5), and 0.011 (Day 7).

Article Snippet: The cultures were then incubated at 37°C in a 5% CO2 atmosphere for 12 h. For those groups receiving blocking antibody treatments, the media also included 100 μg/mL of IgG (I‐1195, Leinco Technologies, St. Louis, MO) or β1‐integrin antibody (AIIB2, Developmental Studies Hybridoma Bank, Iowa City, IA).

Techniques: Migration, RNA sequencing, Control, Functional Assay, Knock-Out, Western Blot, Expressing, Single Cell

Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Timeline and schematic showing the blocking of cell adhesion to nECM using an integrin β1 (ITGB1) function perturbing antibody for 1 hour pre-embedding and for 7 consecutive days . b. Representative fluorescent images of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (scale bar = 100 μm). c. Quantification of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (low-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 30 ROIs, N = 3; high-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 27 ROIs, N = 3). d. Representative fluorescent images of Sox9 immunofluorescence (dashed line outlines nuclei) of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (scale bar = 10 μm). e. Quantification of Sox9 nucleus-to-cytoplasm (NC) ratio of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (low-Ctrl: n = 138 cells, N = 4; low-ITGB1i: n = 133 cells, N = 4; high-Ctrl: n = 124 cells, N = 4; high-ITGBi: n = 138 cells, N = 4). a-e. N = number of independent experiments ****p < 0.0001, *p < 0.05, error bar = standard deviation, ns: no significant difference by two-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: For perturbation studies, media was supplemented with either an anti-CD44 antibody (DSHB, H4C4, 2.5 μg/mL, day 0-3) or anti-integrin β1 antibody (anti-ITGB1, DSHB, AIIB2, 2.5 μg/mL, day 0-7).

Techniques: Blocking Assay, Cell Culture, Inhibition, Modification, Immunofluorescence, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: HDBECs (2×10 5 ) were incubated for 24 h with recombinant TF (0, 0.5 and 2 U/ml) or PAR2-AP (SLIGKV; 20 µM), or with recombinant TF (0.5 U/ml) that was pre-incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Cells were also pre-incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to addition of TF. The cells were harvested after 24 h, into separate aliquots (1×10 5 cells). Total RNA was isolated from one group and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) Inhibitor of CDK p16 INKa mRNA (n=5), and (B) the relative amounts of p16 INKa protein (n=3) calculated from (C) the western blots of p16 INKa protein (using a goat anti-human antibody) and against GAPDH. TF, tissue factor; PAR2, protease-activated receptor 2.

Article Snippet: In other experiments, the cells were pre-incubated at 37°C for 60 min with AIIB2 antibody (20 μg/ml; cat. no. AIIB2-c; Merck KGaA) to block β1-integrin signalling, or with SAM11 antibody (20 μg/ml; cat. no. sc-13504; Santa Cruz Biotechnology, Inc.) to block PAR2 activation prior to the addition of TF.

Techniques: Incubation, Recombinant, Isolation, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: HDBECs (2×10 5 ) were incubated for 24 h with recombinant TF (0, 0.5 and 2 U/ml) or PAR2-AP (SLIGKV; 20 µM), or with recombinant TF (0.5 U/ml) that was pre-incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Groups of cells were also pre-incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to the addition of TF. The cells were harvested after 24 h, into separate aliquots (1×10 5 cells). Total RNA was isolated from one group and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) CDK interacting protein/Wildtype p53-activated fragment p21 CIP1/WAF1 mRNA (n=5), and (B) the relative amounts of p21 CIP1/WAF1 protein (n=3) calculated from (C) the western blots of p21 CIP1/WAF1 protein, (using a mouse anti-human antibody) and against GAPDH. TF, tissue factor; PAR2, protease-activated receptor 2.

Article Snippet: In other experiments, the cells were pre-incubated at 37°C for 60 min with AIIB2 antibody (20 μg/ml; cat. no. AIIB2-c; Merck KGaA) to block β1-integrin signalling, or with SAM11 antibody (20 μg/ml; cat. no. sc-13504; Santa Cruz Biotechnology, Inc.) to block PAR2 activation prior to the addition of TF.

Techniques: Incubation, Recombinant, Isolation, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Examination of the influence of TF on cell cycle indicators. (A) Groups of HDBECs (1×10 5 ) were transfected with the pGL3-promoter vector containing the E2F-1 enhancer sequence. The cells were then treated for 24 h with recombinant TF (0, 0.5 and 2 U/ml), combinations of TF (0.5 U/ml) with the shown antibodies, or with PAR2-AP (SLIGKV; 20 µM) and the luciferase activity was measured within 24 h (n=3). (B) Groups of cells (5×10 4 ) were seeded in 96-well plates and treated with recombinant TF (0, 0.5 and 2 U/ml) for 24 h. The cells were then fixed and probed with an anti-phospho-Thr821/826 human retinoblastoma protein antibody (1:1,000 v/v) for 1 h. The samples were then incubated with an HRP-conjugated donkey anti-goat IgG diluted 1:3,000 v/v) for 1 h, developed with a TMB substrate and the absorptions determined at 450 nm (n=3). (C) Groups of cells (1×10 5 ) were incubated for 24 h with TF (0, 0.5 and 2 U/ml) or PAR2-AP (20 µM), or with recombinant TF (0.5 U/ml) that was pre-incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Groups of cells were also pre-incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to addition of recombinant TF. The cells were harvested after 24 h, total RNA was isolated and the amount of cyclin D1 mRNA determined against that of β-actin (n=4). (D) Groups of cells (1×10 5 ) were treated as aforementioned and cell numbers were determined using crystal violet staining (n=3). TF, tissue factor; PAR2, protease-activated receptor 2; E2F-1; Early region 2 binding factor.

Article Snippet: In other experiments, the cells were pre-incubated at 37°C for 60 min with AIIB2 antibody (20 μg/ml; cat. no. AIIB2-c; Merck KGaA) to block β1-integrin signalling, or with SAM11 antibody (20 μg/ml; cat. no. sc-13504; Santa Cruz Biotechnology, Inc.) to block PAR2 activation prior to the addition of TF.

Techniques: Transfection, Plasmid Preparation, Sequencing, Recombinant, Luciferase, Activity Assay, Incubation, Isolation, Staining, Binding Assay