Journal: Cell communication and signaling : CCS

Article Title: Peritoneal cavity-derived small extracellular vesicles from aged tumor-naïve hosts promote ovarian cancer adhesion and invasion.

doi: 10.1186/s12964-025-02273-1

Figure Lengend Snippet: Fig. 6 Blocking sEV-associated proteins abrogates the enhanced meso-mimetic adhesion observed with aged sEVs. sEVs (5 × 107) purified from perito neal lavage obtained from aged hosts or control (PBS) were incubated with function-blocking antibodies directed against (A) β1 integrin (2 µg), (B) CA125 (MUC16, 1 µg) or (C) LYN kinase (1 µg) in a total volume of 200 ul for 3 h prior to adding to OvCa cells for 24 h. The meso-mimetic adhesion assay was then performed as described in Fig. 3. (D) sEVs (5 × 107) purified from peritoneal lavage obtained from aged hosts or control (PBS) were incubated with the Lyn kinase inhibitor TL0259 (0.1 nM) for 3 h prior to adding to OvCa cells for 24 h. The meso-mimetic adhesion assay was then performed as described in Fig. 3. Assays were performed in triplicate. Data were analyzed using Kruskal-Wallis test and Dunn’s multi-comparison test

Article Snippet: Gels were transferred to a polyvinylidene difluoride membrane (ImmobilonP, Millipore) using a Bio-Rad Trans-Blot SD Semi-Dry Transfer Cell device, and blocked in 5% milk in TBST buffer (25 mmol/l Tris pH 7.5, 150 mmol/l NaCl, 0.1% Tween 20) for 1 h at room temperature (RT), then were incubated overnight with antibodies to CD9, CD63, CD81, TSG101, Annexin A5, Integrin β1, integrin α3, filaggrin, transglutaminase2, Lyn, Mhc1 (Santa Cruz Biotechnology), Integrin α2 (Advanced Cellular Biology), and MUC16/CA125 (Dako) at a 1:100 dilution in 5% milk in TBST at 4 °C with gentle rocking.

Techniques: Blocking Assay, Purification, Control, Incubation, Cell Adhesion Assay, Comparison

Journal: Science Advances

Article Title: Phenotypic discovery and therapeutic evaluation of an ITGA3B1 -targeting antibody-drug conjugate for bladder cancer

doi: 10.1126/sciadv.ady0041

Figure Lengend Snippet: ( A ) ELISA assessing the binding specificity of 2E7 scFv-Fc binding to the ITGA3B1 heterodimer and its individual subunits ( ITGA3 and ITGB1 ), as well as to the structurally related integrin complex ITGA6B4 and its subunits ( ITGA6 and ITGB4 ). HuIgG and BSA were included as negative controls. Data represent the mean ± SD from three technical replicates. A 450 , absorbance at 450 nm. ( B ) Dose-dependent binding of 2E7 scFv-Fc to immobilized ITGA3 , ITGB1 , and ITGA3B1 , as determined by ELISA. ( C ) Flow cytometry analysis of HEK293 cells transiently transfected with ITGA3 , ITGB1 , or both. ( D ) Flow cytometry of MDA-MB-231 cells following siRNA knockdown of ITGA3 , ITGB1 , or both ITGA3 and ITGB1 . ( E ) SPR sensorgrams showing 2E7 scFv-Fc binding to recombinant ITGA3B1 in the presence of either divalent cations or EDTA. Data shown are representative of three independent experiments.

Article Snippet: Target-specific siRNAs against ITGA3 (sc-35684) and ITGB1 (sc-35674), along with a nontargeting control siRNA (sc-37007), were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Transfection, Knockdown, Recombinant

Journal: Science Advances

Article Title: Phenotypic discovery and therapeutic evaluation of an ITGA3B1 -targeting antibody-drug conjugate for bladder cancer

doi: 10.1126/sciadv.ady0041

Figure Lengend Snippet: ( A ) ITGA3 mRNA expression in normal bladder tissues ( n = 18) and primary bladder tumors ( n = 408) based on TCGA data. ( B ) ITGA3 expression across consensus molecular subtypes of bladder cancer from five aggregated clinical cohorts. ( C ) Representative multiplex immunofluorescence images of TMA cores stained for ITGA3 (green), ITGB1 (red), and nuclei [DAPI (4′,6-diamidino-2-phenylindole), blue] of primary bladder carcinomas and normal tissues. Scale bars, 100 μm. ( D to F ) Quantification of ITGA3 expression in tumor versus normal tissues: fluorescence intensity (D), percentage of ITGA3 -positive cells (E), and H -scores (F). **** P < 0.0001.

Article Snippet: Target-specific siRNAs against ITGA3 (sc-35684) and ITGB1 (sc-35674), along with a nontargeting control siRNA (sc-37007), were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Expressing, Multiplex Assay, Immunofluorescence, Staining, Fluorescence

Journal: Nature Communications

Article Title: Control cell migration by engineering integrin ligand assembly

doi: 10.1038/s41467-022-32686-2

Figure Lengend Snippet: a Schematic illustration of precise control of integrin ligand presentation on nanofilaments via peptide assembly. b TEM images of nanofilaments obtained via molecular self-assembly and co-assembly of FFFIKLLI (100 μM) and FFF at various ratios, and the estimated molecular packing structures. IKLLI motif is presented in blue and FFF motif is presented in pink. The scale bars represent 200 nm. Three independent experiments were performed. c Zoom-in SEM images (false color) of HuH-7 cell edge and apical membrane after 3-day incubations. FFFIKLLI was maintained at a concentration of 100 μM. The cell body is highlighted in pink, while the nanofilaments are highlighted in blue. The scale bars represent 300 nm.

Article Snippet: Integrin β1 shRNA plasmid (#sc-29375-SH), Integrin α3 shRNA plasmid (#sc-35684-SH), Integrin α6 shRNA plasmid (#sc-43129-SH), and control shRNA plasmid-A (#sc-108060) were purchased from Santa Cruz Biotechnology for knockdown of the target integrins.

Techniques: Control, Membrane, Concentration Assay

Journal: Nature Communications

Article Title: Control cell migration by engineering integrin ligand assembly

doi: 10.1038/s41467-022-32686-2

Figure Lengend Snippet: a Time-lapse series showing actin cytoskeleton (grey) and paxillin (green) in HuH-7 cells expressing mRuby-Lifeact-7 and mGFP-paxillin upon the treatment of FFFIKLLI (100 μM) for 12 hr. Scale bars represent 2 μm. Three independent experiments were performed. b F-actin phalloidin staining (magenta), integrin β1 (cyan) and paxillin (yellow) immunofluorescence in HuH-7 cell after 12 h treatment of FFFIKLLI (100 μM). Scale bar represents 2 μm. c Fluorescence intensity distribution profile of integrin β1, paxillin, and actin cytoskeleton along the yellow line on merged image of b . d 12 h time course Rac1 activity of HuH-7 cells with or without the treatment of FFFIKLLI (100 μM). Rac1 activity was measured by FRET. n = 6 cells (Ctrl) or 5 cells (FFFIKLLI). Symbols represent the mean FRET/CFP emission ratio ± s.d. e Representative FRET/CFP ratio images of HuH-7 cells expressing RaichuEV-Rac1 with or without the treatment of FFFIKLLI (100 μM) at the indicated time points and coded according to a pseudo-color scale, which ranges from yellow to purple with an increase in Rac1 activity. Scale bars represent 20 μm. Source numerical data are available in source data.

Article Snippet: Integrin β1 shRNA plasmid (#sc-29375-SH), Integrin α3 shRNA plasmid (#sc-35684-SH), Integrin α6 shRNA plasmid (#sc-43129-SH), and control shRNA plasmid-A (#sc-108060) were purchased from Santa Cruz Biotechnology for knockdown of the target integrins.

Techniques: Expressing, Staining, Immunofluorescence, Fluorescence, Activity Assay

Journal: Frontiers in Oncology

Article Title: Inter and intra-tumor heterogeneity of paediatric type diffuse high-grade gliomas revealed by single-cell mass cytometry

doi: 10.3389/fonc.2022.1016343

Figure Lengend Snippet: Summary of the 15 antibodies used for the mass cytometry analysis.

Article Snippet: Anti-Human CD49c , 161Dy , Fluidigm , 3161016B , .

Techniques: Mass Cytometry

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: ( A ) CD46 (green), PNA lectin (red). ( B ) β1 integrin (green), PNA lectin (red). The first column represents a schematic illustration of ( A ) CD46, ( B ) β1 integrin localization in the acrosome intact sperm (line I) and in sperm during the AR progress. ( A ) CD46 detection in intact acrosomal membranes (line I), and the residual outer acrosomal membrane (line II) (see the green arrow), the inner acrosomal membrane begins to emerge (see the red arrow). CD46 relocation progress during the AR is seen across the apical equatorial segment towards the whole equatorial segment and the whole sperm head. ( B ) β1 integrin is relocated across the apical equatorial segment towards the whole equatorial segment and the whole sperm head. In contrast to CD46, the residual acrosome cap and the apical equatorial segments were not detected. Scale bar represents 2 μm.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Membrane

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: Individual bars denote the percentage distribution of CD46 and β1 integrin staining patterns among individual times of capacitation and induced AR. Error bars denote SEM. AC – Acrosome Cap, rAC – residual Acrosome Cap, aES – apical Equatorial Segment, ES – Equatorial Segment, WSH – Whole Sperm Head. C – time of the capacitation, AR – time of the induced acrosome reaction.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Staining

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: ( A ) Coloured lines with error bars represent the relative fluorescent intensities of β1 integrin (red) and CD46 (green) among the individual segments of the sperm head in 10 sperm with an intact acrosome. Horizontal coloured lines represent the arithmetic means of the fluorescent intensities for β1 integrin (red) and CD46 (green). ( B ) Coloured lines with error bars represent the relative fluorescent intensities of β1 integrin (red) and CD46 (green) among the individual segments of the sperm head in 10 acrosome reacted sperm. ( C ) Statistical comparison of the relative fluorescent intensities of β1 integrin (red) and CD46 (green) among individual segments of the sperm head between the acrosome-intact and acrosome-reacted sperm. Horizontal coloured lines represent the arithmetic means of the fluorescent intensities for β1 integrin (red) and CD46 (green). Error bars denote SEM. AC – Acrosome Cap, ES – Equatorial Segment, PAR – Post-Acrosomal Region. p value equal or lower than 0.05 was considered to be significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: Interactions of CD46 and β1 integrin in C57BL/6 spermatozoa determined by Duolink proximity ligation assay. ( A ) freshly released sperm, ( B ) sperm during the induced AR. Positive control (α tubulin/ β tubulin), negative control (β1 integrin/ α tubulin). The small pictures in the corners represent the immunofluorescent dual staining of CD46 and β1 integrin, CD46 (green), β1 integrin (red). Scale bar represents 4 μm.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Proximity Ligation Assay, Positive Control, Negative Control, Staining

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: ( A ) (line I) CD46 (green) is locked over the acrosome vesicle and β1 integrin (red) is confined to both the plasma and the outer acrosomal membrane with prominent labelling of the perforatorium (see the red arrow); (line II) the onset of AR with the membrane vesiculation is visible, as well as the visibility of CD46 and β1 integrin within the acrosome and plasma membranes (see green and red arrows); (line III) the loss of the acrosome including the CD46 and β1 integrin signal is visible. Relocation of CD46 and β1 integrin over the equatorial segment in patchy clusters can be recognised; (line IV) CD46 is localized through the inner acrosome membrane (see the green arrow), the equatorial segment and over the post-acrosome region. β1 integrin shows a similar pattern to CD46 and also remains localized in the perforatorium (see the red arrow). ( B ) (line I) α6 (green) and β1 (red) integrins occupy different regions of the intact sperm head, except the same localization in the plasma membrane over the acrosome and the hook. α6 integrin is continues to be spread over the equatorial segment, even prior to AR (see the green arrows). β1 integrin is further present in the outer acrosomal membrane (when compared with the CD46 dual staining in AI) and perforatorium (see the red arrows). (line II) The α3 (red) integrin pattern is clearly different to the α6 (green) subunit, but remarkably similar to the one of β1 (red, line I) in the acrosome region. α3 is expressed on in the outer acrosomal membrane and the plasma membrane over the acrosome. (line III) The differences in the localization of the α3 (red) integrin and CD46 (green) are visible. The α3 integrin subunit is detectable on the plasma (see also for detail) and the outer acrosomal membrane, when CD46 is defined strictly to the acrosomal membranes only. DAPI (blue); Scale bar represents 1 μm.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Clinical Proteomics, Membrane, Staining

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: (line I and III) Actin (red) fills the apical acrosome and equatorial segment of an intact sperm head, it copies the plasma membrane and overlays the CD46 (green, line I) labelled acrosome in a thin line pattern (see red arrow). (line III) β1 integrin and actin display a similar localization in the apical acrosome cap region (plasma and acrosomal membrane). The perforatorium is filled with actin, but clearly marked with the β1 integrin. (line II and IV) Actin (red) is confined to the equatorial and postacrosomal segment in the acrosome reacted sperm. CD46 (green, line II) and β1 (green, line IV) relocation progress during the AR is visible. DAPI (blue); Scale bar represents 1 μm.

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Clinical Proteomics, Membrane

Journal: Scientific Reports

Article Title: Characterization of CD46 and β1 integrin dynamics during sperm acrosome reaction

doi: 10.1038/srep33714

Figure Lengend Snippet: ( A ) SIM data show the localization of CD46 (green) on the inner and outer acrosomal membrane and β1 integrin (red) on the plasma and outer acrosomal and plasma membrane of the acrosomal area. Scale bar represents 1 μm. ( B ) SIM super-resolution image analysed by Huygens software, showing the colocalization area (yellow) of selected proteins in the outer acrosomal membrane. The colocalization map is based on Pearson’s correlation coefficient. Scale bar represents 1 μm. DAPI (blue).

Article Snippet: The nitrocellulose membranes were incubated with the primary antibody for 1.5 h, washed six times for 5 min with a wash solution (0.05% Tween-20 in PBS) and incubated with a peroxidase-conjugated secondary antibody for 1 h. Proteins were identified as follows; CD46: rat monoclonal antibody (MM10, Hycult Biotech), 1:100 followed by donkey anti rat IgG antibody conjugated to HRP (170 5046, Bio-Rad), 1:10000; β1 integrin: rabbit polyclonal antibody (M-106: sc-8978 Santa Cruz Biotechnology) 1:20, followed by a peroxidase goat anti rabbit IgG (170–6515 BioRad), 1:3000; α3 integrin: primary rabbit polyclonal antibody (H-43): sc-28665 Santa Cruz Biotechnology diluted 1:20 and peroxidase goat anti rabbit IgG secondary antibody conjugated to HRP (170–6515 BioRad), 1:3000; α6 integrin: mouse monoclonal antibody (F-6): sc-374057 Santa Cruz Biotechnology 1:20, followed by a goat anti mouse IgG conjugated to HRP (170–6516 BioRad) 1:5000.

Techniques: Membrane, Clinical Proteomics, Software

Journal: Scientific Reports

Article Title: Development and application of two novel monoclonal antibodies against overexpressed CD26 and integrin α3 in human pancreatic cancer

doi: 10.1038/s41598-019-57287-w

Figure Lengend Snippet: Immunoprecipitation and immunodetection by Western blot of ( A ) integrin α3 and ( B ) CD26 antigen with novel mAbs KU44.22B and KU44.13A using lysates from CaOV-3 ovarian cancer cells and AsPC-1 pancreatic cancer cells respectively. Left panel: Immunoprecipitation was performed with novel mAbs ( A ) KU44.22B and ( B ) KU44.13A (5 µg) using sheep anti-mouse dynabeads. Protein bands around ~140 KDa and ~ 260KDa were immunoprecipitated with mAb KU44.22B (A; left panel) and ~110 KDa by mAb KU44.13A (B; left panel) respectively and stained with SimplyBlue™ SafeStain. The ~50/25 KDa bands represent heavy and light chains of the anti-mouse antibody. *( B ) left panel corresponds to a cropped gel; vertically sliced images of juxtaposed lanes that were non-adjacent in the gel have a clear separation delineating the boundary between the gels. Middle panel: Integrin α3 and CD26 antigen were immunoprecipitated with mAbs ( A ) KU44.22B and ( B ) KU44.13A (5 µg) respectively, and probed with the same antibody (30 µg/ml). Target antigens were not immunodetected with either of the mAbs. Right panel: Integrin α3 and CD26 antigen were immunoprecipitated with mAbs ( A ) KU44.22B and ( B ) KU44.13A respectively (5 µg) or commercial anti-integrin α3 and anti-CD26 antibodies (2 µg) and immunodetected with commercial mAbs sc-374242 and ab89398 as described in Methods. Immunodetection of target antigens immunoprecipitated by novel mAbs and probed with commercial mAbs confirmed the target identity. MW: molecular weight marker.

Article Snippet: In conclusion, in this study, we reported the production of two novel monoclonal antibodies against integrin α3 and CD26 on human pancreatic cancer cells, using the liver metastasis pancreatic cancer cell line CFPAC-1 as immunogen.

Techniques: Immunoprecipitation, Immunodetection, Western Blot, Staining, Molecular Weight, Marker

Journal: Scientific Reports

Article Title: Development and application of two novel monoclonal antibodies against overexpressed CD26 and integrin α3 in human pancreatic cancer

doi: 10.1038/s41598-019-57287-w

Figure Lengend Snippet: Identification of proteins recognised by novel mAbs KU44.13A and KU44.22B by mass spectrometry.

Article Snippet: In conclusion, in this study, we reported the production of two novel monoclonal antibodies against integrin α3 and CD26 on human pancreatic cancer cells, using the liver metastasis pancreatic cancer cell line CFPAC-1 as immunogen.

Techniques:

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Integrin expression in clone C cells. (A) Expression patterns of integrin β1, αv, α6, and α1 in confluent monolayers of clone C cells seeded at LD and HD. Integrin staining is depicted in red, and nuclear staining with Sytox is shown in green. Integrin α3 staining was also observed by immunostaining but not depicted here. (B) Integrin expression in clone C cells was investigated by basolateral biotinylation followed by immunoprecipitation with integrin antibodies as indicated and Western blotted with anti-streptavidin antibodies (for αv and α3 in the top panel and all lanes in the bottom panel). In addition, Western blots were performed on biotin-streptavidin–purified basolateral membranes from HD cells with integrin α1, α5, and α6. (C) Western blots of cell lysates from LD and HD cells using the polyclonal antibody directed against extracellular domain of rabbit integrin β1 (left). (Right) Western blot using the same antibody in the presence of 600 ng/ml recombinant peptide immunogen. (D) Flow cytometry analysis of clone C cells with various integrin antibodies as indicated in the labels.

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques: Expressing, Staining, Immunostaining, Immunoprecipitation, Western Blot, Purification, Recombinant, Flow Cytometry

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Integrin β1 binds to hensin and integrin αV and gets tyrosine phosphorylated in high density cells. (A) In the first panel, 140 μg of DTSSP–cross-linked cell protein from LD and HD phenotypes were immunoprecipitated with anti–integrin β1 CSAT antibodies, and the blots were probed with guinea pig anti-hensin antibody. In control experiment, membrane containing integrin β1 immunoprecipitates described in the first panel was stripped and reprobed with a different anti–integrin β1 antibody MAB1965 (second panel). Chemically cross-linked cell lysates from LD and HD cells were immunoprecipitated with goat anti-hensin/DMBT1 antibody and blotted with anti–integrin β1 MAB1965 antibody (third panel). Control experiment in which the same samples were probed with guinea pig anti-hensin antibody (fourth panel). (B) Tyrosine phosphorylation of integrin β1. (Left) Anti-phosphotyrosine Western blot of integrin β1 immunoprecipitates (described in A) from LD and HD. (Middle) Control Western blot with integrin β1 antibody. (Right) A streptavidin Western blot of biotinylated basolateral membrane proteins (from HD) that were immunoprecipitated with anti–integrin β1 antibody. (C) Interaction of integrin β1 with integrin α subunits was examined by Western blotting of integrin β1 immunoprecipitates with antibodies to integrin αv, α6, and α1 (left lane in first three panels) and α3 and α5 (last two lanes). Interaction of hensin with integrin α subunits was examined by Western blotting of hensin immunoprecipitates with antibodies to integrin αv, α6, and α1 (right lane in first three panels) from HD cells. (D) Interaction of integrin αv with integrin β subunits was examined by Western blotting of integrin αv immunoprecipitates with integrin β1 (first lane), integrin β3 (second lane), and integrin β5 (third lane) antibodies.

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques: Immunoprecipitation, Western Blot

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Effect of anti-integrin antibodies on cell height and cross-sectional area a

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques:

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Role of integrin αv and β1 in terminal differentiation of clone C epithelia. (A) Clone C cells cultured at high density in media containing integrin αv and β1 function-blocking antibodies or integrin α1 function-blocking antibody. Apical and basal actin stained with phalloidin (red) and imaged by confocal microscopy showing en face and XZ optical sections. (B) HD cells cultured with or without integrin function-blocking antibodies as indicated in the labels were examined for their ability to endocytose fluorescein-dextran. Fluorescein-dextran in green and nuclear stain (Sytox red) in red. (C, top) Clone C cells seeded at low density and allowed to form confluent monolayers, when examined by F-Actin staining, have stress fibers on the basal surface and very little apical actin staining (left); however, when the same experiment is carried out in the presence of integrin β1–activating antibody P4G11, apical actin staining was observed with a marked difference in stress fiber staining on the basal surface. (Bottom) LD cells cultured in the absence or presence of integrin β1–activating antibody P4G11 were examined for their ability to endocytose green fluorescence (FITC) dextran. Nucleus was visualized with Sytox Red dye.

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques: Cell Culture, Blocking Assay, Staining, Confocal Microscopy, Fluorescence

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Integrins αv and β1 mediate ECM deposition of hensin. (A) ECM hensin staining: Clone C cells seeded at high density were cultured with various function-blocking integrin antibodies and exposed to anti-hensin antibodies (followed by rhodamine secondary antibodies) before fixation or permeabilization. Nuclear staining is shown in green. (B) Control experiment to examine intracellular hensin: Clone C cells seeded at high density were cultured with function-blocking integrin antibodies and exposed to anti-hensin antibodies after fixation and permeabilization. (C) Hensin secretion in basolateral medium: Equal amounts of combined basolateral medium from HD cells cultured in the presence and absence of integrin αv–and β1–blocking antibodies collected for 3 d was precleared with agarose beads, immunoprecipitated with anti-hensin (DMBT1) antibodies, and probed with anti-hensin (SRCR6/7) antibodies. (D) Clone C cells seeded at low density and cultured in the absence or presence of activating antibodies to integrin β1 and stained with hensin antibodies before (top) and after (bottom) permeabilization. (E) ECM was extracted from clone C cells seeded at identical low densities but cultured in the absence or presence of integrin β1–activating antibodies for 5 d. The ECM samples were then probed with Western blotting with guinea pig anti-hensin antibody.

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques: Staining, Cell Culture, Blocking Assay, Immunoprecipitation, Western Blot

Journal:

Article Title: Role of Integrins in the Assembly and Function of Hensin in Intercalated Cells

doi: 10.1681/ASN.2007070737

Figure Lengend Snippet: Integrin α6 is a candidate for transmitting signals from ECM hensin to the cell interior. Effect of function-blocking α6 integrin antibodies on clone c cells seeded at low density on preformed hensin matrix and cultured for 5 d. Monolayers were then stained with Phalloidin (red) and Sytox (green), and the combined projection of all confocal sections is presented in these panels. XZ section represents a typical cross-section through the monolayer.

Article Snippet: In addition, antibodies against integrin β4 (MAB2058, MAB2060; SC-6628-Santa Cruz Biotechnology, Santa Cruz, CA), integrin β3 (MAB1957; cat. no. 553343-BD Pharmingen, San Jose, CA), integrin α2 (MAB1950), integrin α3 (MAB 1952), and integrin α4 (MAB16983) were also used in immunostaining experiments.

Techniques: Blocking Assay, Cell Culture, Staining