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: bioRxiv

Article Title: Thrombospondin-1 Promotes Circuit-Specific Synapse Formation via β1-Integrin

doi: 10.1101/866590

Figure Lengend Snippet: (A) Schematic representation of the experimental design. Purified RGCs isolated from P7 rat retinas are treated with recombinant human TSP1 or TSP2 (150 ng/mL) in the presence of the function blocking antibody against Integrin β1 or control IgG. RGCs were labeled with a ooDSGC marker, CART (blue), and synaptic markers, Bassoon (pre-, green) and PSD95 (post-, red). (B) Representative images of the synapses formed on the RGCs treated with either full length TSP1 or TSP2 in the presence of integrin β1 function blocking antibody or control IgG. Synapses, co-localized synaptic puncta (yellow) of pre- (Bassoon, green) and post-synaptic (PSD95, red) markers, are marked with white arrows. CART (blue) visualizes ooDSGCs. Full images of RGCs are available in the supplementary figure. Quantification of fold increase in the number of co-localized synaptic puncta from the RGCs treated with (C) function blocking antibody or control IgG demonstrates blocking Integrin β1 specifically inhibits TSP1-mediated synaptogenic activity but not TSP2. Fold increase is calculated by normalizing the number of synapses per cell with the number of synapses per cell in CART+ RGCs in GM Only-Control condition (n=30 cells/condition, One-way ANOVA, *** p<0.0001). (D) Schematic representation of the experimental design. Purified RGCs isolated from P7 rat retinas are transfected with the DNA plasmids transcribing ITGB1 and BFP, then treated with synaptogenic fragments, SD1 or SD2 (150 ng/mL). Then the synapses formed onto the RGCs expressing BFP but negative for CART were imaged for synapse quantification. (E) Representative images of the synapses formed on the RGCs expressing ITGB1 and BFP. The RGCs are treated with either SD1 or SD2, then synapses (white arrows) are labeled as co-localized puncta (yellow) of pre- (Bassoon, green) and post-synaptic (PSD95, red) markers. (F) Quantification of fold increase in the number of co-localized synaptic puncta demonstrates that expression of ITGB1 sufficient for TSP1-mediated synapse formation in CART-negative RGCs. (n=20-30 cells/condition, One-way ANOVA, *** p<0.0001).

Article Snippet: Briefly, 125 ng of each DNA construct expressing BFP and ITGB1 (Addgene 51920) and 1.25μl of Lipofectamine LTX was used in each well (in a 24 well plate).

Techniques: Purification, Isolation, Recombinant, Blocking Assay, Labeling, Marker, Activity Assay, Transfection, Expressing

Journal: bioRxiv

Article Title: Thrombospondin-1 Promotes Circuit-Specific Synapse Formation via β1-Integrin

doi: 10.1101/866590

Figure Lengend Snippet: (A) Schematic representation of the transgenic mice breeding strategy. Retinas from the littermate wild-type (WT, Itgb1 +/+) and Integrin β1 cKO (Itgb1 f/f) were collected at P30. (B) Tamoxifen (20 mg/mL) was subcutaneously delivered twice at P9 and P10 (0.6 mg each day) then retinas were collected at P30. (C) Schematic of IPL and cell types that express Cadherin 6 (blue). (D) Representative images of the excitatory synapses in WT and Itgb1 cKO retinas at P30. Cadherin6-positive DS circuit IPL sublayers are visualized by tdTomato (blue, left panels) and excitatory synapses are labeled by VGluT1 (pre-, green) and PSD95 (post-, red). Only the excitatory synapses within tdTomato positive dendrites are shown by masking with tdTomato staining (right panels). The inlets (white boxes) are shown in higher magnification and the co-localized synaptic puncta (merge, yellow) are marked with white arrows. (E) There is no significant change in the number of excitatory synapses within the IPL between control (+/+) and cKO (f/f). (F) Quantification of excitatory synapses within tdTomato-positive dendrites demonstrates significant reduction in synapse number in Itgb1 cKO retinas (n=3 animals per genotype, t-test, *** p<0.0001). (G) Representative images of the inhibitory synapses in WT and Itgb1 cKO retinas at P30. The inhibitory synapses are labeled by Bassoon (pre-, green) and Gephyrin (post-, red)). Only the synapses within tdTomato positive dendrites are shown by masking with tdTomato staining. The inlets (white boxes) are shown in higher magnification and the co-localized synaptic puncta (merge, yellow) are marked with white arrows. (H) There is a significant reduction in the number of inhibitory synapses within the IPL between control (+/+) and cKO (f/f) (n=3 animals per genotype, t-test, *** p<0.0001). (I) Quantification of inhibitory synapses within tdTomato-positive dendrites demonstrates that there is no change in inhibitory synapse number between Itgb1 WT and cKO retinas (n=3 animals per genotype, t-test, n.s., not significant)

Article Snippet: Briefly, 125 ng of each DNA construct expressing BFP and ITGB1 (Addgene 51920) and 1.25μl of Lipofectamine LTX was used in each well (in a 24 well plate).

Techniques: Transgenic Assay, Labeling, Staining

Journal: bioRxiv

Article Title: Integrin beta 1 and mannose receptor 2 are involved in the antifungal activity of bronchial epithelial cells through Aspergillus fumigatus lectin FleA interactions

doi: 10.64898/2026.02.26.708144

Figure Lengend Snippet: Measurement of galactomannan released in BECs transfected with specific ITGB1, MRC2, LAMP1, ITGB1+MRC2 or control siRNA for 72 h and then infected with A. fumigatus conidia for 15 h. Data are expressed as % of control and presented as mean ± SD, n = 3-6 independent experiments. *p < 0.05, **** p < 0.0001 (ANOVA test followed by Bonferroni’s multiple comparison test)

Article Snippet: Reverse transcriptionwas was performed from 1μg RNA using a 2X RT Master Mix on a ProFlex thermocycler (Applied Biosystems, ThermoFischer), with the following program: 25°C for 10 min; 37°C for 2 h and 85°C for 5 min. Quantitative PCR was carried out using 2 μL of cDNA, 5 μL 2× TaqMan mix (Low ROX), 2.5 μL of RNase-free H 2 0 and 0.5 μL of the TaqMan probe: GAPDH (Hs02786624_g1); ITGB1 (Hs01127536_m1); MRC2 (Hs00195862_m1); LAMP1 (Hs00931461_m1); LAMB3 (Hs00165078_m1); LAMC2 (Hs01043717_m1) (Applied Biosystem).

Techniques: Transfection, Control, Infection, Comparison

Journal: bioRxiv

Article Title: Integrin beta 1 and mannose receptor 2 are involved in the antifungal activity of bronchial epithelial cells through Aspergillus fumigatus lectin FleA interactions

doi: 10.64898/2026.02.26.708144

Figure Lengend Snippet: (A ) Representative sensorgram representing binding of ITGB1 (analyte) to immobilized biotinylated FleA (ligand) in SPR kinetic titration serie for concentrations of 1.85, 5.5, 16.6 and 50 nM (left) and the steady-state affinity fit (right) where raw data are represented as points with the corresponding fitted curve colored for each experiment. ( B ) Representative sensorgram representing binding of ITGB1 (analyte) to immobilized biotinylated FleA (ligand) in BLI kinetic titration serie for concentrations of 15.6, 31.3, 62.5, 125, 250, 500 and 1000 nM (left) and the mean steady-state affinity fit with standard deviation on the duplicate (right). Sensorgrams for the duplicate BLI kinetic titration after subtraction of the reference representing binding of FleA (analyte) to immobilized His tagged ITGB1( C ) or MRC2 ( D ) for concentrations of 3.9, 15.65, 65.5, 250, 1000 and 1500 nM (left) and mean the steady-state affinity fit with standard deviation of the duplicate (right).

Article Snippet: Reverse transcriptionwas was performed from 1μg RNA using a 2X RT Master Mix on a ProFlex thermocycler (Applied Biosystems, ThermoFischer), with the following program: 25°C for 10 min; 37°C for 2 h and 85°C for 5 min. Quantitative PCR was carried out using 2 μL of cDNA, 5 μL 2× TaqMan mix (Low ROX), 2.5 μL of RNase-free H 2 0 and 0.5 μL of the TaqMan probe: GAPDH (Hs02786624_g1); ITGB1 (Hs01127536_m1); MRC2 (Hs00195862_m1); LAMP1 (Hs00931461_m1); LAMB3 (Hs00165078_m1); LAMC2 (Hs01043717_m1) (Applied Biosystem).

Techniques: Binding Assay, Titration, Standard Deviation

Journal: bioRxiv

Article Title: Integrin beta 1 and mannose receptor 2 are involved in the antifungal activity of bronchial epithelial cells through Aspergillus fumigatus lectin FleA interactions

doi: 10.64898/2026.02.26.708144

Figure Lengend Snippet: BECs stimulated with FITC FleA (green) for 15 min, 30 min, 2 h and 4 h or non-stimulated (NS), fixed and stained for ITGB1 (pink) (A) , MRC2 (pink) (B) and LAMP1 (pink) (C). The merge fluorescence between FleA and ITGB1 or MRC2 or LAMP1 is visualized in white. Quantification of white spots (white arrows) visualizing overlay fluorescence (merge) of MRC2 and FleA (D) and LAMP1 and FleA (E) . Data are expressed as number of white puncta per cell and presented as mean ± SD, n=3 independent experiments. *p < 0.05, ** p < 0.01, *** p < 0.001 (ANOVA test followed by Bonferroni’s multiple comparison test)

Article Snippet: Reverse transcriptionwas was performed from 1μg RNA using a 2X RT Master Mix on a ProFlex thermocycler (Applied Biosystems, ThermoFischer), with the following program: 25°C for 10 min; 37°C for 2 h and 85°C for 5 min. Quantitative PCR was carried out using 2 μL of cDNA, 5 μL 2× TaqMan mix (Low ROX), 2.5 μL of RNase-free H 2 0 and 0.5 μL of the TaqMan probe: GAPDH (Hs02786624_g1); ITGB1 (Hs01127536_m1); MRC2 (Hs00195862_m1); LAMP1 (Hs00931461_m1); LAMB3 (Hs00165078_m1); LAMC2 (Hs01043717_m1) (Applied Biosystem).

Techniques: Staining, Fluorescence, Comparison

Journal: bioRxiv

Article Title: Integrin beta 1 and mannose receptor 2 are involved in the antifungal activity of bronchial epithelial cells through Aspergillus fumigatus lectin FleA interactions

doi: 10.64898/2026.02.26.708144

Figure Lengend Snippet: Proximity ligation assay permitting detection of protein-protein interactions in situ (<40 nm distance) at endogenous protein levels was performed in BECs stimulated by FleA for 15 min, 30 min 1 hour and 4 h or not stimulated (NS). Specific ITGB1, MRC2 and LAMP1 antibodies were then linked covalently to specific DNA primers. A hybridization step followed by a PCR amplification with fluorescent probes then permits visualization of ITGB1-MRC2 (A) , ITGB1-LAMP1 (B) , MRC2-LAMP1 (C) interactions as red spots of proximity by fluorescence microscopy. Quantification of red spots visualizing protein-protein interactions is expressed as arbitrary units and presented as mean ± SD, n=3 independent experiments. *p < 0.05 (ANOVA test followed by Bonferroni’s multiple comparison test) (D) .

Article Snippet: Reverse transcriptionwas was performed from 1μg RNA using a 2X RT Master Mix on a ProFlex thermocycler (Applied Biosystems, ThermoFischer), with the following program: 25°C for 10 min; 37°C for 2 h and 85°C for 5 min. Quantitative PCR was carried out using 2 μL of cDNA, 5 μL 2× TaqMan mix (Low ROX), 2.5 μL of RNase-free H 2 0 and 0.5 μL of the TaqMan probe: GAPDH (Hs02786624_g1); ITGB1 (Hs01127536_m1); MRC2 (Hs00195862_m1); LAMP1 (Hs00931461_m1); LAMB3 (Hs00165078_m1); LAMC2 (Hs01043717_m1) (Applied Biosystem).

Techniques: Proximity Ligation Assay, Protein-Protein interactions, In Situ, Hybridization, Amplification, Fluorescence, Microscopy, Comparison

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: Nature Communications

Article Title: Single-cell and spatial transcriptome analyses reveal tumor heterogeneity and immune remodeling involved in pituitary neuroendocrine tumor progression

doi: 10.1038/s41467-025-60028-5

Figure Lengend Snippet: a Prevalence of each immune stroma cluster, estimated by Ro/e score. b −d Proportions of patients with different invasion ( b ), relapse ( c ) and size ( d ) statuses comparing TAM_SPP1-enriched ( n = 5) versus TAM_SPP1-depleted ( n = 25) groups. P -values were determined by two-sided chi-square test. e Patient proportions with varying invasion statuses, using bulk sequencing data from Zhang 2022 ( n = 80 (SPP1 high) and 114 (SPP1 low)) and Zhang 2024 ( n = 16 (SPP1 high) and 16 (SPP1 low)). p-values calculated by two-sided chi-square test. f Bar plot showing the proportion of SPP1+ TAMs in invasive ( n = 14) and non-invasive ( n = 33) PitNETs, as determined by multiplex immunofluorescence staining data. p -value was calculated using the two-sided wilcoxon signed-rank test. Boxes span the interquartile range (IQR) and whiskers extend to points that lie within 1.5 IQRs of the lower and upper quartile. Center line is drawn at the median. g Bar plot of top 5 ligand-receptor pairs with the highest interaction probabilities between invasive and noninvasive PitNETs. h Chord diagrams illustrating interactions between TAM subtypes and tumor clusters via SPP1-ITGAV/ITGB1 ligand-receptor pair, with line width indicating interaction intensity. i Signature scores of MAPK pathways (top) and PI3K-AKT pathways (bottom) for invasive ( n = 44,607 cells) and non-invasive ( n = 103,032 cells) tumor clusters. p-values were determined using the two-sided wilcoxon signed-rank test. Boxes span the interquartile range (IQR) and whiskers extend to points that lie within 1.5 IQRs of the lower and upper quartile. Center line is drawn at the median. j Spatial gene expression patterns of SPP1 , ITGAV , and ITGB1 in samples SCA5 (invasive tumor) and SCA1 (non-invasive tumor). k Representative multiplex immunofluorescence staining of human PitNET tissue. DAPI (blue), CD68 (yellow), SPP1 (orange), ITGB1 (red), ITGAV (green), and tumor marker TPIT (magenta) are shown in individual and merged channels ( n = 3 biological replicates). Scela bar, 100 µm. Source data are provided as a Source Data file for ( a − i ).

Article Snippet: The relative markers CD68 (PA014, abcarta, RTU), SPP1 (ab214050, Abcam, 1:2000), αSMA (19245S, CST, 1:200), ITGAV (AF7308, Beyotime, 1:400), ITGB1 (E-AB-10403, Elabscience, 1:400), FN1 (40932, SAB, 1:300), MYL9 (E-AB-12582, Elabscience, 1:100), ADCYAP1 (FNab10094, FineTest, 1:1500), ANXA1 (AF5114, Beyotime, 1:1000), ASPM (BDPN1665, biodragon, 1:150), BRCA2 (E-AB-40288, Elabscience, 1:1000), CASC5 ( PAB31361 , bioswamp, 1:1000), PENK (BD-PT5829, biodragon, 1:600), Ki67 (ZM-0166, ZSGB, RTU), tumor markers: ACTH (ZM-0004, ZSGB, RTU), PIT-1 (ZM-0208, ZSGB, RTU), SF1 ( GT231702 , Genetech, RTU), and TPIT (ZM-0318, ZSGB, RTU) were evaluated via IHC.

Techniques: Sequencing, Multiplex Assay, Immunofluorescence, Staining, Gene Expression, Marker

Journal: Nature Communications

Article Title: Single-cell and spatial transcriptome analyses reveal tumor heterogeneity and immune remodeling involved in pituitary neuroendocrine tumor progression

doi: 10.1038/s41467-025-60028-5

Figure Lengend Snippet: a Western blot analysis of ITGB1 expression in GH3, AtT-20, and TtT/GF cell lines. b Semi-quantitative analysis of ITGB1 in GH3, AtT-20, and TtT/GF cell lines ( n = 3 biological replicates). c CCK-8 cell viability assay of GH3, AtT-20, and TtT/GF cell lines under specified treatments ( n = 3 biological replicates). d Transwell invasion assay of GH3, AtT-20, and TtT/GF cell lines under specified treatments ( n = 3 biological replicates). e Quantitative analysis of Transwell invasion assay in GH3, AtT-20, and TtT/GF cell lines under specified treatments ( n = 3 biological replicates). Scale bar, 100 µm. Two-sided unpaired t -test was performed for ( b − e ). Data are presented as mean values +/- SD for ( b − e ). Source data are provided as a Source Data file for ( a − e ).

Article Snippet: The relative markers CD68 (PA014, abcarta, RTU), SPP1 (ab214050, Abcam, 1:2000), αSMA (19245S, CST, 1:200), ITGAV (AF7308, Beyotime, 1:400), ITGB1 (E-AB-10403, Elabscience, 1:400), FN1 (40932, SAB, 1:300), MYL9 (E-AB-12582, Elabscience, 1:100), ADCYAP1 (FNab10094, FineTest, 1:1500), ANXA1 (AF5114, Beyotime, 1:1000), ASPM (BDPN1665, biodragon, 1:150), BRCA2 (E-AB-40288, Elabscience, 1:1000), CASC5 ( PAB31361 , bioswamp, 1:1000), PENK (BD-PT5829, biodragon, 1:600), Ki67 (ZM-0166, ZSGB, RTU), tumor markers: ACTH (ZM-0004, ZSGB, RTU), PIT-1 (ZM-0208, ZSGB, RTU), SF1 ( GT231702 , Genetech, RTU), and TPIT (ZM-0318, ZSGB, RTU) were evaluated via IHC.

Techniques: Western Blot, Expressing, CCK-8 Assay, Viability Assay, Transwell Invasion Assay