Journal: Sensors and Actuators B: Chemical

Article Title: Extracellular matrix protein microarray-based biosensor with single cell resolution: Integrin profiling and characterization of cell-biomaterial interactions

doi: 10.1016/j.snb.2019.126954

Figure Lengend Snippet: Fig. 1. Protein microarray biosensor development. (A) Scheme of the

Article Snippet: A suspension of 106 cells was dyed for 10 min at 4 oC in the dark with human CD29-PEVIO770 antibodies for β1 detection (Miltenyi Biotec, Cat. No: 130-101-281) and human CD51/CD61-APC antibodies for αvβ3 determination (Miltenyi Biotec, Cat. No: 130-103-745) following the manufacturer’s instructions.

Techniques: Microarray

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 5. Expression changes of key GBM proteins and their response to FN1 Variants in TBMN. Immunofluorescence experiments assessed the expression of Laminin α5β2, COL4A3/4/5 and Integrin β1 in the GBM. The findings revealed non-homogeneous linear structure expression of these key GBM proteins in the presence of FN1 variants, indicating their potential involvement in the pathogenesis of TBMN. HC = healthy control.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Expressing, Immunofluorescence, Control

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 6. Abnormal co-localization of FN1 variants with Integrin β1 and its impact on other GBM components. Immunofluorescence studies investigated the rela tionship between FN1 variants and the expression patterns of GBM components. The findings reveal abnormal co-localization of FN1 variants with Integrin β1, accompanied by reduced co-localization of other GBM components. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Immunofluorescence, Expressing

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 7. Competitive Binding of FN1 Variants to Integrin β1. (A and B) Duolink proximity ligation assay and Co-IP were employed to analyze the protein interaction between FN1 variants and Integrin β1. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Binding Assay, Proximity Ligation Assay, Co-Immunoprecipitation Assay

Journal: Oncology Reports

Article Title: MicroRNA-3653 inhibits the growth and metastasis of hepatocellular carcinoma by inhibiting ITGB1

doi: 10.3892/or.2019.6971

Figure Lengend Snippet: ITGB1 is a direct target of miR-3653 in HCC. (A) miR-3653 was found to contain the complementary sequences mediating the binding between miR-3653 and ITGB1 3′-UTR. (B) Overexpression of miR-3653 decreased the luciferase activity of the wild-type (wt) ITGB1 3′-UTR but had no obvious influence on that of the mutated (mt) ITGB1 3′-UTR. Knockdown of miR-3653 significantly increased the luciferase activity of the wt 3′-UTR of ITGB1 but had no obvious influence on that of the mt 3′-UTR of ITGB1. (C) Overexpression of miR-3653 significantly decreased the mRNA level of ITGB1 in HCCLM3 cells. (D) Overexpression of miR-3653 significantly decreased the protein level of ITGB1 in HCCLM3 cells. (E) Knockdown of miR-3653 significantly increased the mRNA level of ITGB1 in Hep3B cells. (F) Knockdown of miR-3653 significantly increased the protein level of ITGB1 in Hep3B cells. (G) Immunohistochemical (IHC) staining for ITGB1 in HCC tissues with low miR-3653 levels and those with high miR-3653 levels. Scale bar, 100 µm. HCC, hepatocellular carcinoma; ITGB1; integrin-β1. *P<0.05.

Article Snippet: ITGB1 vector (1.5 µg/ml; cat. no. 51920) and the empty vector (1.5 µg/ml; cat. no. 52535) were obtained from Addgene and co-transfected with miR-3653 mimic or non-targeting control into HCCLM3 cells: HCCLM3 cells co-transfected with non-targeting control (product no. HMC0002) and control vector (cat. no. 52535), HCCLM3 cells transfected with miR-3653 mimic (product no. HMI0001-HMI2785) and control vector (cat. no. 52535), and HCCLM3 cells transfected with miR-3653 mimic (product no. HMI0001-HMI2785) and ITGB1 vector (cat. no. 51920).

Techniques: Binding Assay, Over Expression, Luciferase, Activity Assay, Immunohistochemical staining, Immunohistochemistry

Journal: Oncology Reports

Article Title: MicroRNA-3653 inhibits the growth and metastasis of hepatocellular carcinoma by inhibiting ITGB1

doi: 10.3892/or.2019.6971

Figure Lengend Snippet: ITGB1 mediates the biological functions of miR-3653 in HCC cells. (A) HCCLM3 cells were transfected with control vector or ITGB1 vector. qRT-PCR showed that the ITGB1 vector effectively increased ITGB1 mRNA in HCCLM3 cells. (B) HCCLM3 cells overexpressing miR-3653 were transfected with the control vector or ITGB1 vector. Transfection of ITGB1 vector restored ITGB1 expression in HCCLM3 cells overexpressing miR-3653, and led to decreased E-cadherin and increased N-cadherin. (C and D) Restoration of ITGB1 reversed the inhibitory effects of miR-3653 overexpression on the cell viability and proliferation of HCCLM3 cells. (E) Restoration of ITGB1 reversed the inhibitory effects of miR-3653 overexpression on the migration and invasion of HCCLM3 cells. Magnification, ×40. HCC, hepatocellular carcinoma; ITGB1; integrin-β1. *P<0.05.

Article Snippet: ITGB1 vector (1.5 µg/ml; cat. no. 51920) and the empty vector (1.5 µg/ml; cat. no. 52535) were obtained from Addgene and co-transfected with miR-3653 mimic or non-targeting control into HCCLM3 cells: HCCLM3 cells co-transfected with non-targeting control (product no. HMC0002) and control vector (cat. no. 52535), HCCLM3 cells transfected with miR-3653 mimic (product no. HMI0001-HMI2785) and control vector (cat. no. 52535), and HCCLM3 cells transfected with miR-3653 mimic (product no. HMI0001-HMI2785) and ITGB1 vector (cat. no. 51920).

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Expressing, Over Expression, Migration

Journal: Cells

Article Title: Methylglyoxal Impairs the Pro-Angiogenic Ability of Mouse Adipose-Derived Stem Cells (mADSCs) via a Senescence-Associated Mechanism

doi: 10.3390/cells12131741

Figure Lengend Snippet: mADSCs characterization. ( a ) Representative photographs (5× magnification of an optical microscope, scale bar 500 µm) of mADSCs isolated from the subcutaneous adipose tissue of C57bl6 mice cultured in selection medium for 4 passages. ( b ) Accumulated cell number and ( c ) population doubling time (PTD) per hours of mADSCs from p3 to p11. Once 90% confluence was reached, cells were counted by an automated cell counter. Dots in line graphs represent the mean ± SEM values. ( d – i ) Representative histograms of the expression of CD44, CD29, CD90.2, Sca-1, CD45 and CD31 surface markers analyzed by FACS.

Article Snippet: Then, the cells were divided into 4 aliquots: (i) stained with APC-labeled anti-CD29 (Miltenyi Biotec 130-119-166), FITC-labeled anti-CD45 (Miltenyi Biotec 130-110-796) and PE-labeled anti-CD90.2 (Miltenyi Biotec 130-120-897) in 100 μL of PBS; (ii) PE-labeled anti-CD44 and APC-labeled anti-CD31 antibodies in 100 μL of PBS; (iii) APC-labeled anti-Sca-1 (Miltenyi Biotec 130-123-848) in 100 μL of PBS; or (iv) used as negative control.

Techniques: Microscopy, Isolation, Cell Culture, Selection, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Single-cell RNA sequencing of endometrium uncovers dynamic characteristics and dysregulation of perivascular CD9 + SUSD2 + cells in thin endometrium

doi: 10.1186/s13287-025-04658-y

Figure Lengend Snippet: Collagen deposition surrounding perivascular CD9⁺SUSD2⁺ cells is closely associated with thin endometrium. A CellChat analysis showing the number of incoming signaling interactions from other cell types to perivascular CD9⁺SUSD2⁺ cells in normal versus thin endometrium samples. B Expression patterns of collagen signaling-related genes in normal and thin endometrium. C Western blotting analysis of the COL1A1, ITGB1, α-SMA, and GAPDH in normal and thin endometrium ( n = 6 per group). D The relative protein levels of CD9, SUSD2, and ITGB1 were analyzed between normal and thin endometrium. E Immunofluorescence co-staining of CD9, SUSD2, ITGB1, α-SMA, COL1A1, and DAPI in proliferative and thin endometrial tissues ( n = 10 per group). F - H Quantitative analyses of the percentage of COL1A1 in stromal cells ( F ), ITGB1 ( G ), and α-SMA ( H ) in CD9⁺SUSD2⁺ cells in proliferative and thin endometrial tissues. Ctrl-Pro, normal proliferative endometrium; Thin-Pro, thin proliferative endometrium. Data are presented as mean ± SD. * P < 0.05; ** P < 0.01

Article Snippet: The membranes were incubated with primary antibodies against COL1A1 (1:1500, Cat# PA5-29569, Thermo Fisher, USA), ITGB1 (1:1500, Cat# RA011880A0HU, Cusabio, China), α-SMA (1:2000, Cat# 19245T, CST, USA), or GAPDH (1:4000, Cat# #97166, CST, USA) at 4 °C overnight.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining