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: Developmental cell

Article Title: Global versus local matrix remodeling drives rotational versus invasive collective migration of epithelial cells.

doi: 10.1016/j.devcel.2024.11.021

Figure Lengend Snippet: Figure 5. ICM but not RCM is dependent on MMP-mediated matrix remodeling (A and B) Frequencies of ductal-like and acinus-like morphologies at 7 days in vehicle- and MMPi-treated (A) MDA-MB-231 (n = 217 multicellular structures and n = 192 multicellular structures for vehicle and MMP-treated conditions, respectively) and (B) SK-HEP-1 cell lines (n = 207 multicellular structures and n = 220 multicellular structures for vehicle and MMP-treated conditions, respectively). (C and D) Average angular speed per cellular track of vehicle- and MMPi-treated (C) MDA-MB-231 (n = 18 individual cells, n = 9 cell clusters per treatment condition) and (D) SK-HEP-1 cells in 2-cell clusters (n = 8 individual cells, n = 4 cell clusters per treatment condition). (E and F) Cell persistence ratio per cellular track for vehicle- and MMPi-treated (E) MDA-MB-231 (n = 18 individual cells, n = 9 cell clusters per treatment condition) and (F) SK-HEP-1 cells in 2- cell clusters (n = 8 individual cells, n = 4 cell clusters per treatment condition). (G) Time-lapse images of circular 2-cell clusters of SK-HEP-1 MMPi-treated cells reveals persistent rotational migration. Scale bar, 10 mm. (H and I) Quantification of (H) pericellular matrix-remodeling intensity and (I) % pericellular matrix remodeling, as measured by DQ-collagen I. (J) Quantification of MMP-dependent remodeling, using the collagen 3=4 antibody specific to MMP-cleaved collagen I (n = 15 cells per condition were analyzed from three independent biological replicates). (K) Representative fluorescent micrographs show matrix remodeling of vehicle-treated invasive and rotational and MMPi-treated rotational MDA-MB-231 cells, as measured by DQ-collagen I and collagen 3=4 antibody. Scale bar, 10 mm. (L) Representative fluorescent micrographs show MT1-MMP and ITGb1 signal distribution in vehicle-treated invasive and rotational and MMPi-treated rotational MDA-MB-231 cells. Scale bar, 25 mm. (M and N) Quantification of (M) MT1-MMP and (N) ITGb1 signal distribution along pericellular membranes of vehicle-treated invasive and rotational and MMPi-treated rotational MDA-MB-231 cells, as quantified by MT1-MMP signal distribution skewness (n = 14–15 cells per condition were analyzed from three independent biological replicates). (O–Q) Quantification of (O) maximum protrusion length, (P) average protrusion lifetime, and (Q) protrusion formation rate, as measured by mCherry-LifeAct. (R–T) Quantification of (R) average maximum bead displacement, (S) average % moving beads, and (T) bead speed, as measured by traction force beads. For graphs (H, I, and O–T) n = 9–10 cells were analyzed per condition. All error bars represent standard deviation from the mean. Statistical significance is indicated as ns, not significant, (*) p % 0.05, (**) p % 0.01, (***) p % 0.001, and (****) p % 0.0001.

Article Snippet: CRISPR-Cas9 mediated gene suppression of ITGb1 Genetic knockdown of ITGb1 was performed as described previously.15 In brief, small guide RNAs targeting the ITGb1 gene were cloned into lentiCRISPR v2 plasmid (Addgene #52961) following instructions from Feng Zhang, the plasmid supplier.

Techniques: Migration, Standard Deviation

Journal: Developmental cell

Article Title: Global versus local matrix remodeling drives rotational versus invasive collective migration of epithelial cells.

doi: 10.1016/j.devcel.2024.11.021

Figure Lengend Snippet: Figure 6. RCM relies on ITGb1 (A) Western blot assessment of ITGb1 depletion in MDA-MB-231 cell line. (B) Frequencies of ductal-like and acinus-like morphologies at 7 days in WT and crITGb1 MDA-MB-231 cells (n = 139 multicellular structures and n = 141 multicellular structures for WT and crITGb1 cell lines, respectively). (C) Polar plots of rounded 2-cell clusters of MDA-MB-231 crITGb1 cells (n = 6 individual cells, n = 3 cell clusters), and color indicates cluster pair. (D) Representative fluorescent micrographs show matrix-remodeling localization at 48 h of rounded WT and crITGb1 MDA-MB-231 cells, as measured by DQ-collagen I. Scale bar, 10 mm. (E) Quantification of pericellular matrix-remodeling intensity, as measured by DQ-collagen I (n = 15 cells for all WT conditions, n = 25 for all crITGb1 conditions). (F) Western blot assessment MT1-MMP expression in MDA-MB-231 WT and crITGb1 cells. Values on blot indicate quantified relative MT1-MMP expression. (G) Quantification of MMP-dependent remodeling, using the collagen 3=4 antibody specific to MMP-cleaved collagen I. n = 15 cells for all WT conditions, n = 25 for all crITGb1 conditions. All error bars represent standard deviation from the mean. (H and I) Histology of whole-mount mammary glands from 6.5-week-old mice treated daily with (H) vehicle or (I) GM-6001 broad-spectrum MMP inhibitor from 3.5 weeks old reveals increased lateral budding in GM6001-treated mammary glands. Scale bar, 1 mm. Image reproduced with permission: ª2003 Wiseman et al.44 Originally published in Journal of Cell Biology. https://doi.org/10.1083/jcb. 200302090 as Figures 2a and 2b. (J) Histology of mammary tissue from wild-type control (left column) and Col1a1tm1Jae (right column) mice showing increased prevalence of spheroid structures (arrows) in Col1a1tm1Jae mammary tissue. Collagen surrounding the mammary duct is detected with H&E, trichrome, and picrosirius red staining. Images taken from Provenzano et al.46 Figures 3a and 3c and Supplementary Figures 2a and 2b, which was published in BMC Medicine in 2006 with a Creative Commons CC BY license (https://doi.org/10.1186/1741-7015-4-38). Statistical significance is indicated as ns, not significant, (*) p % 0.05, (**) p % 0.01, (***) p % 0.001, and (****) p % 0.0001.

Article Snippet: CRISPR-Cas9 mediated gene suppression of ITGb1 Genetic knockdown of ITGb1 was performed as described previously.15 In brief, small guide RNAs targeting the ITGb1 gene were cloned into lentiCRISPR v2 plasmid (Addgene #52961) following instructions from Feng Zhang, the plasmid supplier.

Techniques: Western Blot, Expressing, Standard Deviation, Control, Staining

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: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 1. The effect of GLUT1 on NSCLC cell proliferation and colony formation. (A, B) NSCLC cell proliferation; (C, D) NSCLC cell colony formation. Adding siRNA-GLUT1 (siGLUT1) to A549 cell and GLUT1 expression plasmid (GLUT1) to LK2 cell; the graph in (C) and (D) shows the number of colony formation under different treatments; *, P <0.05 compared with negative control; NC, negative control; si-NC, scramble siRNA for negative control; Error bars, S.D.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Expressing, Plasmid Preparation, Negative Control

Journal: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 2. The effect of GLUT1 on the cell cycle correlated proteins in NSCLC cells. We added siRNA-GLUT1 (siGLUT1) to A549 cell and GLUT1 expression plasmid (GLUT1) to LK2 cell; NC, negative control; si-NC, scramble siRNA for negative control.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Expressing, Plasmid Preparation, Negative Control

Journal: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 3. The effect of GLUT1 on NSCLC cell migration and correlated proteins. (A, B) NSCLC cell migration; (C, D) the expressions of ROCK1, ROCK2 and RhoA. We added siRNA-GLUT1 (siGLUT1) to A549 cell and GLUT1 expression plasmid (GLUT1) to LK2 cell; the graph in (A) and (B) shows the distance of cell migration under different treatments; *, P<0.05 compared with negative control; NC, negative control; si-NC, scramble siRNA for negative control; Error bars, S.D.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Migration, Expressing, Plasmid Preparation, Negative Control

Journal: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 4. The effect of GLUT1 on NSCLC cell invasion and correlated proteins. (A, B) NSCLC cell invasion; (C, D) the expressions of MMP2, proMMP2 and TIMP2. We added siRNA-GLUT1 (siGLUT1) to A549 cell and GLUT1 expression plasmid (GLUT1) to LK2 cell; the graph in (A) and (B) shows the number of invading cells under different treatments; *, P<0.05 compared with negative control; NC, negative control; si-NC, scramble siRNA for negative control; Error bars, S.D.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Expressing, Plasmid Preparation, Negative Control

Journal: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 5. The effect of GLUT1 on the apoptosis in A549 and LK2 cells. We added siRNA-GLUT1 (siGLUT1) to A549 cell (A) and GLUT1 expression plasmid (GLUT1) to LK2 cell (B); NC, negative control; si-NC, scramble siRNA for negative control.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Expressing, Plasmid Preparation, Negative Control

Journal: Journal of Cancer

Article Title: Glucose Transporter 1 Promotes the Malignant Phenotype of Non-Small Cell Lung Cancer through Integrin β1/Src/FAK Signaling.

doi: 10.7150/jca.30772

Figure Lengend Snippet: Figure 6. The effect of GLUT1 on integrin β1/Src/FAK signaling in NSCLC cells. We added siRNA-GLUT1 (siGLUT1) to A549 cell and GLUT1 expression plasmid (GLUT1) to LK2 cell; NC, negative control; si-NC, scramble siRNA for negative control.

Article Snippet: GLUT1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35493), integrin β1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (CA, USA; catalog number: sc-35674), and GLUT1 plasmid were purchased from OriGene (Rockville, USA; catalog number: SC116011).

Techniques: Expressing, Plasmid Preparation, Negative Control