human itgb4  (Santa Cruz Biotechnology)

Journal: Cancers

Article Title: Fibroblasts Promote Resistance to KRAS Silencing in Colorectal Cancer Cells

doi: 10.3390/cancers16142595

Figure Lengend Snippet: List of anti-human antibodies used for flow cytometry.

Article Snippet: CD104 , FITC , REA236 , 130-124-266 , Miltenyi Biotec.

Techniques: Cytometry

Journal: Cancers

Article Title: Fibroblasts Promote Resistance to KRAS Silencing in Colorectal Cancer Cells

doi: 10.3390/cancers16142595

Figure Lengend Snippet: List of anti-human antibodies used for flow cytometry.

Article Snippet: CD104 , FITC , REA236 , 130-124-266 , Miltenyi Biotec.

Techniques: Cytometry

Journal: The American journal of pathology

Article Title: Periostin in Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression by Enhancing Cancer and Stromal Cell Migration.

doi: 10.1016/j.ajpath.2023.12.010

Figure Lengend Snippet: Figure 5 The enhancement of esophageal squamous cell carcinoma cell migration by periostin is mediated via integrin b4. A and B: Integrin subunit beta 4 (ITGB4) and integrin b4 expression levels in TE cells after direct co-culture were compared with those after mono-culture using real-time quantitative PCR (qPCR; A) and Western blot analysis (B). Beta actin (ACTB) was used as a control in the Western blot analysis. CeE: TE-9, TE-10, and TE-15 cells were transfected with siRNA targeting ITGB4 (siITGB4; 20 nmol/L) and negative control siRNA (siNC; 20 nmol/L). ITGB4 knockdown was confirmed using qPCR (C), RT-PCR (D), and Western blot analysis (E). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ACTB were used as a control in RT-PCR and Western blot analysis, respectively. F: Transwell migration assays were performed to confirm the effect of ITGB4 knockdown on the enhanced migration of TE-9, TE-10, and TE-15 cells by adding recombinant human periostin (rhPOSTN; 100 pg/mL). Migrating cells were counted in five random fields in each chamber after 48 hours of incubation. G: TE-9, TE-10, and TE-15 cells transfected with siNC and siITGB4 were treated with rhPOSTN (100 pg/mL), and time-dependent changes in Akt, phosphorylated Akt (p-Akt; Ser473), p-Akt (Thr308), extracellular signal-regulated kinase (Erk), and phosphorylated Erk (p-Erk; Thr202/Tyr204) levels in each cell were then determined using Western blot analysis. ACTB was used as a control. Data are presented as means SEM (A, C, and F). **P < 0.01, ***P < 0.001. N.S., not significant.

Article Snippet: For ITGB4 knockdown, TE-9, TE-10, and TE-15 cells were transfected with siRNA targeting human ITGB4 (20 pmol; sc-35678; Santa Cruz Biotechnology) using Lipofectamine RNAiMAX (Invitrogen) for 48 hours, according to the manufacturer’s instructions.

Techniques: Migration, Expressing, Co-Culture Assay, Real-time Polymerase Chain Reaction, Western Blot, Control, Transfection, Negative Control, Knockdown, Reverse Transcription Polymerase Chain Reaction, Recombinant, Incubation

Journal: The American journal of pathology

Article Title: Periostin in Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression by Enhancing Cancer and Stromal Cell Migration.

doi: 10.1016/j.ajpath.2023.12.010

Figure Lengend Snippet: Figure 9 Schematic diagram of roles of periostin (POSTN) in the esophageal squamous cell carcinoma (ESCC) microenvironment. Direct contact with ESCC cells leads mesenchymal stem cells (MSCs) to become cancer-associated fibroblast (CAF)elike cells, which secrete periostin and activate the Akt and extra- cellular signal-regulated kinase (Erk) pathways via integrin subunit beta 4 (ITGB4) in ESCC cells, promoting their migration. Periostin also promotes MSC and macrophage migration and contributes to the activation of tumor-associated macrophage (TAM)elike macrophage properties. MF, macrophage.

Article Snippet: For ITGB4 knockdown, TE-9, TE-10, and TE-15 cells were transfected with siRNA targeting human ITGB4 (20 pmol; sc-35678; Santa Cruz Biotechnology) using Lipofectamine RNAiMAX (Invitrogen) for 48 hours, according to the manufacturer’s instructions.

Techniques: Migration, Activation Assay

Journal: PLoS ONE

Article Title: Acceleration of Bone Repair in NOD/SCID Mice by Human Monoosteophils, Novel LL-37-Activated Monocytes

doi: 10.1371/journal.pone.0067649

Figure Lengend Snippet: A. Monocytes were incubated in the absence or presence of LL-37 (5 µM), GM-CSF (20 ng/mL), M-CSF (50 ng/mL), or M-CSF and RANKL (both at 25 ng/mL) for 6 days. Surface staining of Integrin α3 and α3β1 were analyzed using flow cytometry . B–C. Monocytes were incubated in the presence of 5 µM LL-37 for different days and proliferative capacity was detected using either ( B ) FITC BrdU/7AAD flow kit or ( C ) Cell proliferation dye eFluor 670. Data shown were from three independent experiments.

Article Snippet: Anti-human integrin β3 (Clone: 25E11), anti-human integrin β4 (Clone: 3E1) and anti-human integrin β5 (Cat#: AB1926-20) were from EHD Millipore (Billerica, MA, USA), PE anti-human CD49a (Integrin α1, clone: TS2/7), FITC anti-human CD49b (Integrin α2, clone: AK-7), Alexa Fluor 488 anti-human CD49c (Integrin α3, Clone ASC-1), PE anti-human CD49d (Integrin α4, clone: 9F10), APC anti-human CD49e (Integrin α5, clone : NK-SAM-1), APC anti-human CD49f (Integrin α6, clone: GoH3), FITC anti-human CD68 (CloneY1/82A) and Alexa Fluor 647 anti-CCR4 (Clone TG6/CCR4) from Biolegend (San Diego, USA), TaqMan array human Pathway Plate from Invitrogen Corp (Grand Island, USA), cell proliferation Dye eFluor670 and eBioscience Fixation/Permeabilization Concentrate and Diluent from eBioscience (San Diego, USA), MMP-7 ELISA Kit from RayBiotech, Inc (Norcross, USA), anti-human p38MAPKα/β/γ (Rabbit), anti-phospho-ERK1/2 (Rabbit), anti-phospho-Smad1/5/8 (Rabbit) and anti-MAPK13 (p38MAPKδ) (Rabbit) from Cell Signaling Technology (Danvers, USA), and EIA kits of Osteopontin, GPNMB/osteoactivin, Cartilage glycoprotein-39 (also called Chitinase 3-like 1), Chitinase 1, MMP-7, and CCL22 from R&D systems (Minneapolis, USA).

Techniques: Incubation, Staining, Flow Cytometry