integrin α5β1 inhibitor  (TargetMol)

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

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

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

doi: 10.1002/advs.202409147

Figure Lengend Snippet: Figure 6. 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: In Vitro Inhibition of SOX18, Integrin α5β1, and MAPK Cascade: The SOX18 inhibitor, Sm4,[92] was purchased from MedChemExpress (Catalog HY-116940), with a working concentration of 10 μmol L−1 for in vitro assays.

Techniques: Expressing, Inhibition, Comparison

Journal: Nature Communications

Article Title: Epidermal growth factor-like domain 7 drives brain lymphatic endothelial cell development through integrin αvβ3

doi: 10.1038/s41467-024-50389-8

Figure Lengend Snippet: a FISH and antibody staining shows ilk is expressed in GFP+ BLECs at 4 dpf (arrowheads). n = 17/18 embryos. b – g Inhibition of ILK by cpd22 can partially rescue the absence of BLECs in the egfl7 mutant at 6 dpf ( b – e ). Quantification of the number of BLECs in the double loops of brain in different treatment groups ( f , n = 10 embryos, two-tailed unpaired t test. Data are represented as mean ± SD). The statistics show the percentage of embryos that have double lymphatic loops, single loops, and none of the loops in the brain ( g , WT, n = 51 embryos, WT + cpd22, n = 67 embryos, egfl7-/- , n = 116 embryos, egfl7-/- + cpd22, n = 127 embryos, χ 2 test).). h – k An egfl7 mutant was crossed with an ilk heterozygote to generate two types of larvae: ilk +/+; egfl7 -/- and ilk +/-; egfl7 -/-, which were then studied for BLECs loop-structures formation. The larvae were classified into three categories based on their phenotype: Double loops, single loops, and none of the loops ( h – j ). The results showed that compared to ilk +/+; egfl7 -/-, the ilk +/-; egfl7 -/- larvae had a decreased percentage of the none of the loops phenotype, and a partially rescued percentage of double loops and single loops phenotype ( k , ilk +/+; egfl7 -/-, n = 49 embryos, ilk +/-; egfl7 -/-, n = 47 embryos, χ2 test). Scale bar, 50 μm.

Article Snippet: The integrin α5β1 inhibitor-ATN-161(Selleck, #262438-43-7) and the integrin αvβ3 inhibitor-Cilengitide (MCE, #HY16141) stock solution of 10 mM in DMSO was used to prepare a working solution-50 μM and treat the embryos from 54 hpf to 5 dpf.

Techniques: Staining, Inhibition, Mutagenesis, Two Tailed Test

Journal: Molecular medicine reports

Article Title: Astaxanthin inhibits integrin α5 expression by suppressing activation of JAK1/STAT3 in Helicobacter pylori‑stimulated gastric epithelial cells.

doi: 10.3892/mmr.2023.13014

Figure Lengend Snippet: Figure 1. ASX inhibits integrin α5 expression in Helicobacter pylori‑stimulated AGS cells. Western blot analysis of integrin α5 and β1 in cells (A) stimulated with H. pylori for the indicated time periods or (B) pretreated with the indicated concentrations of ASX for 3 h, and then stimulated with H. pylori for 6 h. The densitometry data are presented as the mean ± standard error from three immunoblots, and are shown as the relative density of protein bands normalized to β‑actin. *P<0.05 vs. (A) 0 h or (B) ‘None’ (unstimulated cells without ASX treatment). (B) #P<0.05 vs. ‘Control’ (stimulated cells without ASX treatment). ASX, astaxanthin.

Article Snippet: The JAK/STaT inhibitor aG490 (cat. no. T3434; r&d Systems, inc.) and integrin α5β1 antago‐ nist K34c (N‐(2,6‐dimethylbenzoyl)‐O‐[3‐(2‐pyridinylamino) propyl]‐l‐tyrosine; cat. no. 5114; r&d Systems, inc.) were dissolved in dMSo at 50 mM.

Techniques: Expressing, Western Blot, Control

Journal: Molecular medicine reports

Article Title: Astaxanthin inhibits integrin α5 expression by suppressing activation of JAK1/STAT3 in Helicobacter pylori‑stimulated gastric epithelial cells.

doi: 10.3892/mmr.2023.13014

Figure Lengend Snippet: Figure 4. AG490 and K34C suppress Helicobacter pylori‑induced integrin α5 expression, cell adhesion and migration. (A) Cells were pretreated with 40 µM AG490 for 2 h, and subsequently stimulated with H. pylori for 6 h. Protein levels of integrin α5 and β1 were determined using western blot analysis, with β‑actin as the loading control. (B) Cells were pretreated with 40 µM AG490 or 20 µM K34C for 2 h, and subsequently stimulated with H. pylori for 24 h. Adherent cells were stained and absorbance was detected at 570 nm. Results are presented as the mean ± standard error (the total number for each group was 12). The percentage of adherent cells in the ‘None’ group was set as 100%. (C) Cells were pretreated with 40 µM AG490 or 20 µM K34C for 2 h, and subsequently stimulated with H. pylori for 20 h. The wound healing assay was performed to detect cell migration. Representative images of wounds in AGS cells were captured at baseline (0 h) and at 20 h (magnification, x100). Wound closure was evaluated by measuring the remaining cell‑free area and expressed as a percentage of the initial cell‑free area. The bar graph indicates wound closure rate (%). Results are presented as the mean ± SE (the total number for each group was 12). (D) Cells were pretreated with ASX (5 µM), AG490 (40 µM) and K34C (20 µM), and then stimulated with H. pylori for 6 h. The pretreatment period for ASX was 3 h, while that of AG490 or K34C was 2 h. Protein levels of integrin α5 and β1 were determined using western blot analysis, with β‑actin as the loading control. The densitometry data are presented as the mean ± standard error from three immunoblots, and are shown as the relative density of protein bands normalized to β‑actin. *P<0.05 vs. ‘None’ (unstimulated cells without treatment of AG490, K34C or ASX). #P<0.05 vs. ‘Control’ (H. pylori‑stimulated cells without treatment of AG490, K34C or ASX). ASX, astaxanthin.

Article Snippet: The JAK/STaT inhibitor aG490 (cat. no. T3434; r&d Systems, inc.) and integrin α5β1 antago‐ nist K34c (N‐(2,6‐dimethylbenzoyl)‐O‐[3‐(2‐pyridinylamino) propyl]‐l‐tyrosine; cat. no. 5114; r&d Systems, inc.) were dissolved in dMSo at 50 mM.

Techniques: Expressing, Migration, Western Blot, Control, Staining, Wound Healing Assay