Journal: Stem Cell Research & Therapy

Article Title: Extracellular matrix derived from Wharton’s Jelly-derived mesenchymal stem cells promotes angiogenesis via integrin αVβ3/c-Myc/P300/VEGF

doi: 10.1186/s13287-022-03009-5

Figure Lengend Snippet: Oligonucleotide primers and PCR conditions in RT-qPCR

Article Snippet: FAK inhibitors (PF-573228), antagonist for integrin αVβ3 (HY-100563) and integrin α5β1 (HY-13535A) were obtained by MCE Inc. (USA).

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Extracellular matrix derived from Wharton’s Jelly-derived mesenchymal stem cells promotes angiogenesis via integrin αVβ3/c-Myc/P300/VEGF

doi: 10.1186/s13287-022-03009-5

Figure Lengend Snippet: Integrin αVβ3 mediated the effect of WJ-MSCs ECM on HUVECs. a–e mRNA expressions of integrin α5, αV, β1, β3, and β5 in HUVECs were determined by RT-qPCR. Mean ± S.E.M., n = 5. f, o–s Protein expressions of FAK, p-FAK, P38, p-P38 and VEGF in HUVECs were determined by western blotting. Mean ± S.E.M. n = 3. g – n Matrigel tube formation assay images of cumulative tube length. Bar = 50 μm. HY-100563: integrin αVβ3 specific antagonist. HY-13535A: integrin α5β1 specific antagonist. The P value was calculated using one-way ANOVA and independent samples t -test. * P < 0.05, ** P < 0.01 versus control

Article Snippet: FAK inhibitors (PF-573228), antagonist for integrin αVβ3 (HY-100563) and integrin α5β1 (HY-13535A) were obtained by MCE Inc. (USA).

Techniques: Quantitative RT-PCR, Western Blot, Tube Formation Assay, Control

Journal: Stem Cell Research & Therapy

Article Title: Extracellular matrix derived from Wharton’s Jelly-derived mesenchymal stem cells promotes angiogenesis via integrin αVβ3/c-Myc/P300/VEGF

doi: 10.1186/s13287-022-03009-5

Figure Lengend Snippet: Integrin αVβ3 mediated the activation of FAK/P38 signaling pathway induced by WJ-MSCs ECM in HUVECs. a–e Transwell for HUVECs migration assay and quantitative analysis of migrated HUVECs numbers. Bar = 50 μm. Mean ± S.E.M., n = 3. f–j Immunofluorescently stained for VEGF of HUVECs (magnification: × 200). Mean ± S.E.M., n = 3. HY-100563: Integrin αVβ3 specific antagonist. k–m Protein expressions of FAK, p-FAK, P38 and p-P38 in HUVECs were determined by western blotting. Mean ± S.E.M. n = 3. The P value was calculated using one-way ANOVA and independent samples t -test. * P < 0.05, ** P < 0.01 versus control

Article Snippet: FAK inhibitors (PF-573228), antagonist for integrin αVβ3 (HY-100563) and integrin α5β1 (HY-13535A) were obtained by MCE Inc. (USA).

Techniques: Activation Assay, Migration, Staining, Western Blot, Control

Journal: Stem Cell Research & Therapy

Article Title: Extracellular matrix derived from Wharton’s Jelly-derived mesenchymal stem cells promotes angiogenesis via integrin αVβ3/c-Myc/P300/VEGF

doi: 10.1186/s13287-022-03009-5

Figure Lengend Snippet: ECM derived from WJ-MSCs promoted angiogenesis via integrin αVβ3/c-Myc/P300/VEGF. ECM: Extracellular matrix. αVβ3: integrin αVβ3. HUVEC: human umbilical vein endothelial cell. c-Myc: cellular-Myc. VEGF: vascular endothelial growth factor

Article Snippet: FAK inhibitors (PF-573228), antagonist for integrin αVβ3 (HY-100563) and integrin α5β1 (HY-13535A) were obtained by MCE Inc. (USA).

Techniques: Derivative Assay

Journal: American Journal of Cancer Research

Article Title: Desmoglein-2 promotes the proliferation and invasion of lung adenocarcinoma cells by inhibiting anoikis through the activation of the integrin beta-1/focal adhesion kinase signaling pathway

doi: 10.62347/DWTV3761

Figure Lengend Snippet: DSG2 activates the integrin β1/FAK signaling pathway. A. After transfecting the DSG2-overexpressing plasmid into NCI-H3255 cells and interfering RNA into NCI-H2347 cells for 48 h, a Western blot was used to detect the changes in the relative expression levels of proteins related to the integrin β1/FAK signaling pathway, including integrin β1, p-integrin β1, FAK, p-FAK, and DSG2 in NCI-H3255 cells and NCI-H2347 cells. B. After constructing a DSG2-knockdown cell line in NCI-H2347 cells, Pyrintegrin was added, and incubation continued for 24 h. Western blot was used to detect the changes in the relative expression levels of proteins related to the integrin β1/FAK signaling pathway, including integrin β1, p-integrin β1, FAK, p-FAK, and DSG2 in NCI-H2347 cells. C. After successfully establishing a DSG2-knockdown cell line in NCI-H2347 cells, ATN-161 (an inhibitor of the integrin β1/FAK signaling pathway) was added. Subsequently, the expression levels of proteins, including integrin β1, p-integrin β1, FAK, p-FAK, and DSG2, were detected through Western blot experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, “ns” represents P > 0.05.

Article Snippet: ATN-161 (HY-13535; MedChemExpress, China), an integrin β1/FAK signaling pathway inhibitor, was applied in experiments dividing cells into vector, oe-DSG2, and oe-DSG2 + ATN-161 groups.

Techniques: Plasmid Preparation, Western Blot, Expressing, Knockdown, Incubation

Journal: American Journal of Cancer Research

Article Title: Desmoglein-2 promotes the proliferation and invasion of lung adenocarcinoma cells by inhibiting anoikis through the activation of the integrin beta-1/focal adhesion kinase signaling pathway

doi: 10.62347/DWTV3761

Figure Lengend Snippet: ATN-161 counteracts the promotive effects of DSG2 overexpression on proliferation and invasion in NCI-H3255 cells and induces anoikis. A. After establishing the DSG2-overexpressing cell line in NCI-H3255 cells, ATN-161 was added, and incubation continued for 24 hours. Changes in cell proliferation capacity were detected using the EdU assay (×200, scale bar: 50 µm). B. The expression level of DSG2 in NCI-H3255 cells was detected by immunofluorescence (×200, scale bar: 50 µm). C. Statistical analysis bar chart for the NCI-H3255 cell experiment. D. Statistical analysis bar chart for NCI-H3255 cell immunofluorescence. E. Changes in the proliferation capacity of NCI-H3255 cells were detected by the CCK-8 assay. F. Changes in the invasive capacity of NCI-H3255 cells were detected by the Transwell assay (×200, scale bar: 50 µm). G. Morphological changes of anoikis were observed in NCI-H3255 cells (×100, scale bar: 100 µm). H. Changes in the proportion of apoptotic NCI-H3255 cells were detected by flow cytometry. I. Changes in the relative expression levels of anoikis-related proteins P53, Bcl-2, Bax, Caspase-3, and Cleaved-caspase-3 in NCI-H3255 cells were detected by Western blot. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, “ns” represents P > 0.05.

Article Snippet: ATN-161 (HY-13535; MedChemExpress, China), an integrin β1/FAK signaling pathway inhibitor, was applied in experiments dividing cells into vector, oe-DSG2, and oe-DSG2 + ATN-161 groups.

Techniques: Over Expression, Incubation, EdU Assay, Expressing, Immunofluorescence, CCK-8 Assay, Transwell Assay, Flow Cytometry, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: The dipeptide prolyl-hydroxyproline promotes cellular homeostasis and lamellipodia-driven motility via active β1-integrin in adult tendon cells

doi: 10.1016/j.jbc.2021.100819

Figure Lengend Snippet: Engagement of β1-integrin and ERK signaling in Pro-Hyp-mediated cell migration. A , left panels , immunofluorescence staining for β1-integrin (clone 9EG7, which recognizes an extracellular epitope of ligand-inducible active β1 : green )/DAPI ( blue ) and F-actin ( red )/DAPI ( blue ). Adult tenocytes were treated with 500 μg/ml Pro-Hyp for 6 h. The scale bars represent 20 μm. Right panels , the numbers of active β1-integrin–containing focal contacts and the thickness of F-actin filaments when cells were treated with 500 μg/ml Pro-Hyp for 6 h. Error bars represent the standard deviation (n = 4; four separate culture experiments). ∗∗ p < 0.01; ∗∗∗ p < 0.001. B , effect of MEK1/2 inhibitor PD98059 or α5β1-integrin antagonist ATN-161 on Pro-Hyp–mediated cell migration in adult tenocytes using Boyden chamber. Cell migration activity is shown relative to the control value of 100% (cells treated with 200 μg/ml Pro-Hyp). Error bars represent the standard deviation (n = 5). ∗∗∗ p < 0.001 (in post hoc analysis). C , effect of ATN-161 on uptake of stable isotopically labeled Pro-Hyp (SI-Pro-Hyp; 200 μg/ml) in adult tenocytes for 60 min at 37 °C. Error bars represent the standard deviation (n = 8). ∗∗∗ p < 0.001. DAPI, 4′,6-diamidino-2-phenylindole; ERK, extracellular signal–regulated kinase; MEK1/2, MAPK kinase 1/2; Pro-Hyp, prolyl-4-hydroxyproline.

Article Snippet: MEK1/2 inhibitor PD98059 (IC 50 = 2 μM) was from Calbiochem, and α5β1 integrin antagonist ATN-161 was from Tocris.

Techniques: Migration, Immunofluorescence, Staining, Standard Deviation, Activity Assay, Control, Labeling

Journal: Advanced Science

Article Title: Intratumoral Collagen Deposition Supports Angiogenesis Suggesting Anti‐angiogenic Therapy in Armored and Cold Tumors

doi: 10.1002/advs.202409147

Figure Lengend Snippet: Collagen upregulates SOX18 expression through the integrin α5β1/MEK/ERK signaling pathway. a) Expression levels of collagen receptors across different cell types in the scRNA‐seq dataset, with ITGB1 and ITGA5 showing the highest positive correlation. b) Spatial charting of ITGA5‐B1 and VWF in the P10_T4 LUAD sample from Marco De Zuani et al.’s study, [ ²⁶ ] alongside ITGA5‐B1 expression in endothelial and non‐endothelial cells in the spatial transcriptomic dataset. Statistical significance was determined using the Chi‐square test. *** P < 0.001. c) Expression levels of SOX18, ITGA5, and ITGB1 in tumor cells (A549 and H1299), HUVECs, and CAFs. d) Enhanced co‐localization of ITGA5 and ITGB1 following collagen stimulation. Scale bar = 5 µm. e) Impact of collagen stimulation and ITGA5‐B1 blockade on the MEK/ERK pathway in HUVEC cells. f) Effects of collagen stimulation and MEK/ERK inhibition on SOX18 expression in HUVEC cells. g) Effects of collagen stimulation, ITGA5‐B1 blockade, and MEK/ERK blockade on the subcellular localization of SOX18 in HUVEC cells. Scale bar = 5 µm. Statistical significance was determined using ANOVA with Tukey's multiple‐comparison test. ns, non‐significance, *** P < 0.001. n = 3 per group. h) Effects of collagen stimulation, ITGA5‐B1 blockade, and MEK/ERK blockade on the expression of SOX18 downstream targets, MMP7 and CXCL12, in HUVEC cells. Statistical significance was determined using ANOVA with Tukey's multiple‐comparison test. ** P < 0.01, *** P < 0.001. n = 3 per group. Abbreviations: ITGB1: integrin beta 1; ITGA5: integrin alpha 5; VWF: von‐Willebrand factor; SOX18: sex determining region Y box 18; CAF: cancer‐associated fibroblast.

Article Snippet: The integrin α5β1 inhibitor, ATN‐161, [ ] was obtained from TargetMol (catalog T10397) at a working concentration of 1 µmol L −1 , and the MAPK inhibitor, U0126, [ ] was purchased from TargetMol (catalog T21332) with a working concentration of 5 µmol L −1 for in vitro assays.

Techniques: Expressing, Inhibition, Comparison