rabbit anti human integrin αv polyclonal antibody  (Proteintech)

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a The graph displays the −log10-transformed RRA scores of genes enriched following infection with two SAFV-3 strains in HeLaN-∆SLC cells, analyzed using the MAGeCK software. The X -axis represents data from the screen using the SAFV-3 JPN08-356 strain, whereas the Y -axis shows results from the screen using the SAFV-3 JPN08-404 strain. The dotted line indicates the significance threshold of RRA = 0.01. Genes that met the criterion of RRA < 0.01 in both screens are highlighted in blue. The size of each dot reflects the combined enrichment across both screens, with larger dots indicating a greater sum of −log10 (RRA scores) from both experiments. b Expression of human integrin αV and integrin β8 in HeLaN-WT, HeLaN-∆AV, HeLaN-∆SLC∆AV, HeLaN-∆B8, and HeLaN-∆SLC∆B8 cells. The cells were stained with anti-integrin αV or anti-integrin αVβ8 antibodies and analyzed by flow cytometry. c HeLaN-WT, HeLaN-∆AV, HeLaN-∆B8, HeLaN-∆SLC∆AV, and HeLaN-∆SLC∆B8 cells were infected with tenfold serial dilutions of SAFV-3 and viable cells were stained with crystal violet to assess infection levels. Images are representative of two independent experiments. d Multi-step growth kinetics of SAFV-3 in HeLaN-∆B8, HeLaN-∆SLC∆B8, and HeLaN-WT cells. The cells were infected with SAFV-3 and incubated for up to 5 days. Data are presented as mean viral titers with s.d. ( n = 3). Statistical significance was determined using the two-sided Welch’s t -test. **, P < 0.01, *, P < 0.05, n.s. not significant. The dotted line indicates the limit of detection. Source data are provided as a Source Data file.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Transformation Assay, Infection, Software, Expressing, Staining, Flow Cytometry, Incubation

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a Western blot analysis of integrin αV (left panel) and integrin β8 (right panel) expression in BHK-21 cells. BHK-21 cells that were lentivirally transduced with either human integrin αV (BHK + human AV) or hamster integrin β8 (BHK + hamster B8) were used as positive controls. The anti-integrin αV antibody cross-reacted with both human and hamster integrin αV. Actin served as the loading control. b Expression of HS, integrin αV, and β8 in BHK-21 derivatives. BHK-21 cells were stained with anti-HS antibody (upper left panel). BHK-21 cells stably expressing human integrin αV and/or β8 (BHK + human AV, BHK + human B8, BHK + human AVB8), as well as the control cells, were stained with anti-integrin αV or anti-integrin αVβ8 antibodies. The cells were analyzed by flow cytometry. c Susceptibility analysis using SAF/UnaG in BHK-21 cells expressing human integrin αV and/or β8. UnaG-positive cells (green, upper panel) and Hoechst-stained nuclei (blue, lower panel) were imaged at 16 h post-infection. Scale bar, 200 μm. The percentage of infected cells was determined by examining at least 1000 cells. Data are representative of two independent experiments. d One-step growth kinetics of SAFV-3 in BHK + human AV, BHK + human B8, BHK + human AVB8, and control cells. The cells were infected with SAFV-3 and incubated for up to 24 h. The dotted line indicates the limit of detection. e Western blot analysis of exogenous integrin β8 expression in BHK-21 cells lentivirally transduced with either mouse or hamster integrin β8. The anti-integrin β8 antibody cross-reacted with both mouse and hamster integrin β8. Actin served as the loading control. f Susceptibility analysis using SAF/UnaG in mouse and hamster integrin β8 expressing BHK-21 cells. UnaG-positive cells (green, upper panel) and Hoechst-stained nuclei (blue, lower panel) were captured at 16 h post-infection. Scale bar, 200 μm. Images are representative of two independent experiments. Data in ( c and d ) represent means with s.d. ( n = 3). Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparison test ( c ) and the two-sided Welch’s t -test ( d ). **, P < 0.01, *, P < 0.05, n.s. not significant. Source data are provided as a Source Data file.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Western Blot, Expressing, Transduction, Control, Staining, Stable Transfection, Flow Cytometry, Infection, Incubation, Comparison

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a Expression of human integrin β subunits (β1, β3, β5, and β6) on the surface of BHK-21 derivatives. BHK-21 cells lentivirally transduced with the respective integrin β subunits were stained with the indicated antibodies and analyzed using flow cytometry. b Susceptibility analysis using SAF/UnaG in human integrin αV and the indicated β subunit expressing BHK-21 cells. UnaG-positive cells (green, upper panel) and nuclei stained with Hoechst (blue, lower panel) were imaged at 16 h post-infection. Scale bar, 200 μm. Images are representative of two independent experiments.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Expressing, Transduction, Staining, Flow Cytometry, Infection

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a Pull-down assay of SAFV-3 using heparin (left panel) and integrin αVβ8 (right panel). Heparin and Fc chimera of extracellular domains of integrin αVβ8, αVβ3, or the signal sequence (ss) of integrin αV (negative control) were prepared as complexes with magnetic beads. These complexes were incubated with SAFV-3, followed by western blot analysis of the bound virus using anti-SAFV-3 antiserum (left and right upper panels). The bottom right panel shows an image of the integrin-Fc complex on magnetic beads used for pulldown, detected using an anti-mouse IgG antibody. b Cell surface attachment assay for SAFV-3. HeLaN-WT, HeLaN-∆SLC, HeLaN-∆B8, HeLaN-∆SLC∆B8, and HeLaN-∆SLC + human AVB8 were incubated with SAFV-3, followed by RT-qPCR analysis of the bound virus. c Expression of human integrin β8 in HeLaN-∆SLC and HeLaN-∆SLC + human AVB8 cells. To compare the expression levels of integrin αVβ8 on the cell surface, HeLaN-∆SLC and HeLaN-∆SLC + human AVB8 cells were stained with anti-integrin αVβ8 antibodies and analyzed by flow cytometry. d , e Inhibition of SAFV-3 attachment to the cell surface by soluble heparin ( d ) or recombinant integrin αVβ8 ( e ). HeLaN-WT cells ( d ) or HeLaN-∆SLC + human AVB8 cells ( e ) were incubated with SAFV-3 pretreated with 1 or 10 μg of soluble heparin or recombinant integrin αVβ8, respectively. Recombinant integrin αVβ3 was the negative control. After incubation at 4 °C for 2 h, bound virus was analyzed using RT-qPCR. f HeLaN-WT, HeLaN-∆SLC, HeLaN-∆B8, HeLaN-∆SLC∆B8, and HeLaN-∆SLC + human AVB8 cells were infected with tenfold serial dilutions of SAFV-3, and viable cells were stained with crystal violet to assess infection levels. All data are representative of two independent experiments. Data in ( b , d , and e ) represent means with s.d. ( n = 3). Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparison test. **, P < 0.01, n.s. not significant. Asterisks directly placed on bars indicate statistically significant differences compared to WT or untreated samples, while asterisks placed on the lines connecting bars denote statistically significant differences between those bars. Source data are provided as a Source Data file. Ag antigen, Fc fragment crystallizable region.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Pull Down Assay, Sequencing, Negative Control, Magnetic Beads, Incubation, Western Blot, Virus, Quantitative RT-PCR, Expressing, Staining, Flow Cytometry, Inhibition, Recombinant, Infection, Comparison

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a Expression of HS and human integrin β8 in BHK-WT, BHK-∆SLC, BHK + human AVB8, BHK-∆SLC + human AVB8, and revertant cells expressing human SLC35B2 (BHK-∆SLC + human AVB8 + SLC). The cells were stained with anti-HS or anti-integrin αVβ8 antibodies and analyzed by flow cytometry. b Cell surface attachment assay for SAFV-3. BHK-WT and BHK-∆SLC cells were incubated with SAFV-3 at 4 °C for 2 h. Viral binding to HS was assessed by RT-qPCR quantification of cell-bound virus ( n = 3). c Susceptibility analysis using SAF/UnaG in BHK + human AVB8, BHK-∆SLC + human AVB8, and BHK-∆SLC + human AVB8 + SLC cells. The cells used in this experiment were sorted to equalize the surface expression levels of integrin αVβ8 between BHK + human AVB8 and BHK-∆SLC + human AVB8 cells. UnaG-positive cells (green) and nuclei stained with Hoechst (blue) were imaged at 16 h post-infection. The percentage of infected cells was determined by examining at least 800 cells/well ( n = 4). Scale bar, 200 μm. d Cell surface attachment assay for SAFV-3 in BHK + human AVB8, BHK-∆SLC + human AVB8, and BHK-∆SLC + human AVB8 + SLC cells ( n = 3). Bar graphs in ( b– d ) are presented as means with s.d. Statistical significance was determined using the two-sided Welch’s t -test ( b ) and a one-way ANOVA with Dunnett’s multiple comparison test ( c , d ). **, P < 0.01, *, P < 0.05. All data are representative of two independent experiments. Source data are provided as a Source Data file.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Expressing, Staining, Flow Cytometry, Incubation, Binding Assay, Quantitative RT-PCR, Virus, Infection, Comparison

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: a Viral infection analysis using integrin β8 mutants. BHK-21 cells expressing the human integrin β8 mutants (∆SDL, Y172N, and I208R) were inoculated with SAFV-3. After 2 days, virus titers were determined using the TCID 50 assay. Human integrin β3 was the negative control. The dotted line indicates the limit of detection. The right panel shows the results of western blot analysis of integrin β8 mutants and integrin β3 expression in BHK-21 cells. b Alignment of the amino acid sequences of puff A on VP2 of SAFV-3 (left panel) and CD loop I on VP1 of SAFV-2 (right panel). RGD-like sequences are highlighted. c Infection blocking assay using RGD peptide. Left panel: HeLaN-∆SLC cells were pretreated with 10 or 100 μg of GRGDS, GRADS, or GRAES peptide at 4 °C for 30 min and then incubated with SAFV-3/UnaG virus for an additional 60 min. Right panel: HeLaN-∆SLC cells were pretreated with 10 or 100 μg of GRGDS, GRLDS, or GRAES peptide for 30 min and then incubated with SAFV-2/UnaG virus for an additional 60 min. GRGDS and GRAES peptides were positive and negative controls, respectively. The number of UnaG-positive cells at 14 h post-infection was counted using ImageJ software. d Schematic illustration of mutagenesis in RGD-like sequence. The mutated amino acid residues are highlighted. e HeLaN-∆SLC∆B8, HeLaN-∆SLC, and HeLaN-∆SLC + human AVB8 cells were infected with tenfold serial dilutions of mutant viruses carrying mutations in the RGD-like sequences, and viable cells were stained with crystal violet. Primary progeny virus produced from BHK cells transfected with infectious RNA (P-0 virus) was used. Bar graphs in ( a and c ) represent means with s.d. ( n = 3 in ( a ); n = 9 peptide-free and n = 3 peptide-added samples in ( c )). Statistical significance was determined using a one-way ANOVA with Dunnett’s multiple comparison test. **, P < 0.01, n.s. not significant. Statistical comparisons in ( c ) were made between peptide-free and peptide-added samples. All data are representative of two independent experiments. Source data are provided as a Source Data file.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Infection, Expressing, Virus, Negative Control, Western Blot, Blocking Assay, Incubation, Software, Mutagenesis, Sequencing, Staining, Produced, Transfection, Comparison

Journal: Nature Communications

Article Title: Saffold virus exploits integrin αvβ8 and sulfated glycosaminoglycans as cooperative attachment receptors for infection

doi: 10.1038/s41467-025-67236-z

Figure Lengend Snippet: Sulfated GAGs and integrin αVβ8 function as interconnected dual receptors for SAFV infection in HeLa-N cells. SAFV can directly bind to either sulfated GAGs or integrin αVβ8, while a portion of viruses bound to sulfated GAGs subsequently interact with integrin αVβ8. In addition, the data suggest the existence of a downstream molecule (factor X) required for an unspecified step in the viral entry process following sulfated GAGs binding.

Article Snippet: To detect of endogenous hamster integrin αV and β8, rabbit anti-human integrin αV polyclonal antibody (27096-1-AP, Proteintech) and rabbit anti-mouse integrin β8 (D1V7M) monoclonal antibody (88300, Cell Signaling Technology) were used, respectively.

Techniques: Infection, Binding Assay

Journal: Nature Communications

Article Title: Galectin-3-integrin α5β1 phase separation disrupted by advanced glycation end-products impairs diabetic wound healing in rodents

doi: 10.1038/s41467-025-62320-w

Figure Lengend Snippet: A Quantifications of HMEC-1 cell migration (left) and tube formation (right) induced by recombinant Gal-3 under an insulin-resistant state (treated with serum-free medium containing 100 nM insulin for 24 h) were shown ( n = 4 biological replicates each group). -, normal state; +, insulin-resistant state. B RT-qPCR analysis of VEGFA , FGF2 , and HGF in HMEC-1 cells treated with the indicated concentration of recombinant Gal-3 for 12 h. Relative expression levels were normalized to ACTB ( n = 3 biological replicates each group). C Heatmap of proteomic abundance (normalized using Z-score) of the top 5 Gal-3-interacting proteins in skin endothelial cells from healthy donors (dataset PXD019909, ProteomeXchange). D HMEC-1 cell migration (left) ( n = 4 biological replicates) and tube formation (right) ( n = 3 biological replicates) induced by recombinant Gal-3 with knockdown of Catenin α-1 or integrin β1. E GST pull-down assays. HMEC-1 cell lysate was incubated with GST or GST-Gal-3 and pulled down with GS beads (left panel); cells were treated with GST or GST-Gal-3 at 4 °C for 1 h, cross-linked, lysed and pulled down with GS beads (right panel). GST served as a negative control. Immunoblot analysis of integrin β1 was shown. F Recombinant Gal-3-induced migration of HMEC-1 cells with integrin β1-targeting shRNAs or non-targeting shRNA (shscr) ( n = 3 biological replicates each group). G Recombinant Gal-3-induced migration of HMEC-1 cells incubated with integrin β1 functional blocking antibody (TDM29, 10 µg/mL) or IgG control (left). Quantifications were shown ( n = 3 biological replicates each group). Scale bar, 500 μm. H Schematic diagram of the α subunit partnering with the integrin β1 subunit created in BioRender. Chen, S. (2025) https://BioRender.com/p10vue6 . Among the 12 α subunits, α3, α5, and α6 subunits were detected by the mass spectrometry analysis in the His-Gal-3 immunoprecipitation assay performed in HMEC-1 cells (see Supplementary Fig. ). I GST pull-down assays. HMEC-1 cell lysate was incubated with GST or GST-Gal-3 and pulled down with GS beads (left panel); cells were treated with GST or GST-Gal-3 at 4 °C for 1 h, cross-linked, lysed and pulled down with GS beads (right panel). GST served as a negative control. Immunoblot analysis of integrin α5, integrin α6 and integrin α3 was shown. J Recombinant Gal-3 induced migration of HMEC-1 cells incubated with integrin α5 functional blocking antibody P1D6 (10 µg/mL) ( n = 3 biological replicates each group). K Recombinant Gal-3-induced migration of HMEC-1 cells that were pre-incubated with integrin α5β1 antagonist ATN-161 (100 nM) for 48 h ( n = 5 biological replicates each group). L , Immunoblot analysis of the phosphorylation of integrin β1 (p-integrin β1, Ser783) in HMEC-1 cells that were incubated with recombinant Gal-3 (10 μg/mL). Relative expression levels were normalized to integrin β1, and quantifications were shown below the blots. M Immunoblot analysis and quantifications of p-integrin β1 in wounds of diabetic mice that i.c . injected with OE-Gal-3 adenovirus or Veh. Relative expression levels were normalized to integrin β1 ( n = 3 biological replicates each group). All statistical data points are represented as means ± SEM. P values were determined by unpaired two-tailed Student’s t -test ( A , B , D , F , G , J , K , M ) or one-way ANOVA with Fisher’s LSD post hoc test ( B , D , F ). Error bars mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source Data file. Exact p values are provided in the Source Data file.

Article Snippet: After blocking with 5% non-fat dry milk in PBST (PBS containing 0.05% Tween-20), blots were incubated overnight at 4°C with the following primary antibodies: CD31 (1:1000, ab281583, Abcam, USA), β-Actin (1:1000, 612657, BD Biosciences, USA), Gal-3 (1:1000, ab76245, Abcam, USA), collagen I (1:1000, GB114197 , Servicebio, China), collagen III (1:1000, EPR17673 , Abcam, USA), AKT (1:1000, 9272 s, cell signaling technology, USA), p-AKT (Ser473) (1:1000, 9271 s, cell signaling technology, USA), FoxO1 (1:1000, 18592-1-AP, Proteintech, China), integrin β1 (1:1000, 610467, BD Biosciences, USA), integrin α3 (1:1000, 66070, Proteintech, China), integrin α5 (1:1000, sc-10729, Santa Cruz, USA), integrin α6 (1:1000, ab181551, Abcam, USA), p-integrin β1 (Y783) (1:1000, ab62337, Abcam, USA), VEGFR2 (1:1000, 2479S, Cell signaling, USA), p-VEGFR2 (Tyr1175) (1:1000, 2478S, Cell signaling technology, USA), EGFR (1:1000, 4267S, Cell signaling, USA), p-EGFR (Tyr1068) (1:1000, 3777S, Cell signaling technology, USA), integrin αv(1:1000, 27096-1-AP, Proteintech, China), FAK (1:1000, 12636-1-AP, Proteintech China), p-FAK (Y397) (1:1000, 611806, BD Biosciences, USA), CD146 (1:1000, 611208, BD Biosciences, USA), GST (1:1000, KM8005, Sungene Biotech, China), His-tag (1:1000, RM1001, Beijing Ray Antibody Biotech, China), α-Tubulin (1:1000, F0063, Selleck, USA).

Techniques: Migration, Recombinant, Quantitative RT-PCR, Concentration Assay, Expressing, Knockdown, Incubation, Negative Control, Western Blot, shRNA, Functional Assay, Blocking Assay, Control, Mass Spectrometry, Immunoprecipitation, Phospho-proteomics, Injection, Two Tailed Test

Journal: Nature Communications

Article Title: Galectin-3-integrin α5β1 phase separation disrupted by advanced glycation end-products impairs diabetic wound healing in rodents

doi: 10.1038/s41467-025-62320-w

Figure Lengend Snippet: A – E STZ-induced diabetic rats were i.c . injected with lgals3 adenovirus or control virus (Veh) after wounding, followed by treatment with integrin β1 functional blocking antibody (Anti-β1) or IgG control (IgG). A Representative images of the wounds (left) and percentage of wound closure (right) ( n = 5). B H&E staining of healed wounds. The distance between the first and second yellow dotted lines represents epidermis thickness (red arrows), and the distance between the second and third yellow dotted lines represents granulation thickness (black arrows). Quantifications of the epidermis and granulation thickness were shown on the lower ( n = 5). Scale bar, 500 µm. C Picrosirius red staining showing COL1 and COL3 in healed wound under polarized light. Quantifications of COL1 area percentage and total COL1 and COL3 area were shown on the right ( n = 5). Scale bar, 50 µm. D Immunohistochemical staining of CD31 that marked microvessels (black arrows) in healed wounds. Quantifications of microvessel count per field were shown on the right ( n = 5). Scale bar, 50 µm. E Immunoblot analysis and quantifications of CD31 in healed wounds ( n = 5). F , G STZ-induced diabetic rats were i.c . injected with shRNA targeting integrin α5 (shα5) or non-targeting shRNA (shscr) 2 weeks before wounding, following i.c . injected with recombinant lgals3 adenovirus (OE-Gal-3) or control virus (Veh). Normal rats (Normal) were set as negative control. F COL1 and COL3 in picrosirius red staining in healed wounds. Quantifications of COL1 area percentage and total COL1 and COL3 area were shown ( n = 5). Scale bar, 50 µm. G Immunohistochemical staining of CD31 (black arrows) in healed wounds. Quantifications of microvessel count per field were shown on the right ( n = 5). Scale bar, 50 µm. H Recombinant Gal-3-induced migration of HMEC-1 cells treated with FAK inhibitor (25 μM), Src-inhibitor (1 μM) or IKK inhibitor (0.5 μM) for 48 h. (Veh, n = 5 biological replicates; FAK inhibitor, n = 3 biological replicates; Src-inhibitor, n = 5 biological replicates; IKK inhibitor, n = 5 biological replicates). I Immunoblot analysis of the phosphorylation of FAK (p-FAK, Y397) in HMEC-1 cells treated with recombinant Gal-3 (10 µg/mL). p-FAK levels were normalized to FAK. Quantifications were shown below the blots. J Recombinant Gal-3-induced tube formation of HMEC-1 cells treated with FAK inhibitor (FAKi, 25 μM), ( n = 4 biological replicates). K Immunoblot analysis (upper) and quantifications (lower) of p-FAK in HMEC-1 cells treated with si-integrin β1 (si-β1) or negative control. p-FAK levels were normalized to FAK ( n = 3 biological replicates). L , M HFD/STZ mice were i.c . injected with recombinant lgals3 adenovirus (OE-Gal-3) or control virus (Veh) after wounding, followed by treatment with FAKi (OE-Gal-3+ FAKi, 15 µM, 100 µL/mouse) or vehicle once every two days. L COL1 and COL3 in picrosirius red staining in healed wounds (left). Quantifications of COL1 area percentage and total COL1 and COL3 area (right). Scale bar, 50 µm. M Immunohistochemical staining and quantifications of CD31 (black arrows) in healed wounds. ( n = 3, two sections per mouse). Scale bar, 50 µm. All statistical data are presented as means ± SEM. P values were determined by unpaired two-tailed Student’s t -test ( A – H , J – M ) or one-way ANOVA with Fisher’s LSD post hoc test ( H ). Error bars mean ± SEM of each group. * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source Data file. Exact p values are provided in the Source Data file.

Article Snippet: After blocking with 5% non-fat dry milk in PBST (PBS containing 0.05% Tween-20), blots were incubated overnight at 4°C with the following primary antibodies: CD31 (1:1000, ab281583, Abcam, USA), β-Actin (1:1000, 612657, BD Biosciences, USA), Gal-3 (1:1000, ab76245, Abcam, USA), collagen I (1:1000, GB114197 , Servicebio, China), collagen III (1:1000, EPR17673 , Abcam, USA), AKT (1:1000, 9272 s, cell signaling technology, USA), p-AKT (Ser473) (1:1000, 9271 s, cell signaling technology, USA), FoxO1 (1:1000, 18592-1-AP, Proteintech, China), integrin β1 (1:1000, 610467, BD Biosciences, USA), integrin α3 (1:1000, 66070, Proteintech, China), integrin α5 (1:1000, sc-10729, Santa Cruz, USA), integrin α6 (1:1000, ab181551, Abcam, USA), p-integrin β1 (Y783) (1:1000, ab62337, Abcam, USA), VEGFR2 (1:1000, 2479S, Cell signaling, USA), p-VEGFR2 (Tyr1175) (1:1000, 2478S, Cell signaling technology, USA), EGFR (1:1000, 4267S, Cell signaling, USA), p-EGFR (Tyr1068) (1:1000, 3777S, Cell signaling technology, USA), integrin αv(1:1000, 27096-1-AP, Proteintech, China), FAK (1:1000, 12636-1-AP, Proteintech China), p-FAK (Y397) (1:1000, 611806, BD Biosciences, USA), CD146 (1:1000, 611208, BD Biosciences, USA), GST (1:1000, KM8005, Sungene Biotech, China), His-tag (1:1000, RM1001, Beijing Ray Antibody Biotech, China), α-Tubulin (1:1000, F0063, Selleck, USA).

Techniques: Injection, Control, Virus, Functional Assay, Blocking Assay, Staining, Immunohistochemical staining, Western Blot, shRNA, Recombinant, Negative Control, Migration, Phospho-proteomics, Two Tailed Test

Journal: Nature Communications

Article Title: Galectin-3-integrin α5β1 phase separation disrupted by advanced glycation end-products impairs diabetic wound healing in rodents

doi: 10.1038/s41467-025-62320-w

Figure Lengend Snippet: A Confocal microscopy of integrin α5 segregation in HMEC-1 cells treated with recombinant Gal-3 (1.65 µM) plus lactose (10 mM). An enlarged view of the boxed region shows clusters on the cell membrane (red arrows). Quantifications of cluster number per cell (left to right, n = 5, 6, 5 fields; total number of cells were 50-90 in each group). Scale bar, 20 µm. B , C Condensates formed with the GFP-Gal-3 (80 µM), GFP-Gal-3 (80 µM) + integrin β1 (400 nM) mixture, GFP-Gal-3 (80 µM) + integrin β1 (400 nM) mixture plus lactose (20 mM), and GFP-Gal-3 (80 µM) + CD146 (400 nM) mixture in PBS (pH 7.4), respectively. B Fluorescence microscopy images of the condensates (red arrows) and quantifications of condensates’ number and diameter in each microscope field ( n = 5 biological replicates). Scale bar, 10 µm. C Solution turbidity for the indicated mixtures measurements by UV-vis spectrophotometry ( n = 3 biological replicates each group). D , E Condensates formed with the GFP-Gal-3 (80 µM), GFP-Gal-3 (80 µM) + integrin α5β1 (400 nM) mixture, GFP-Gal-3 (80 µM) + integrin α5β1 (400 nM) mixture plus lactose (20 mM), and GFP-Gal-3 (80 µM) + CD146 (400 nM) mixture in PBS (pH 7.4). D Fluorescence microscopy images of the condensates and quantifications of condensates’ number, and diameter of each microscope field were shown ( n = 5 biological replicates). Scale bar, 10 µm. E Solution turbidity for the mixtures (left to right, n = 3, 4, 3, 3 biological replicates). F Solution turbidity for the mixtures formed with Gal-3 (0 µM, 10 µM, 20 μM, 40 µM, 80 µM and 100 µM) and integrin α5β1 (400 nM) ( n = 3 biological replicates). G Fluorescence Recovery After Photobleaching (FRAP) analysis of droplets formed with GFP-Gal-3 (80 µM) and integrin β1 (400 nM), integrin α5β1 (400 nM) or CD146 (400 nM), respectively. Representative confocal microscopy images (left) and normalized fluorescence intensity (right) after bleaching were shown ( n = 5, 4, 5 independent measurements, respectively). H Condensates formed with the GFP-Gal-3 (80 µM) and integrin α5β1 (400 nM) in PBS (pH 7.4) had their N-glycans removed by PNGase. Fluorescence microscopy images of the condensates and quantifications of condensates’ number, and diameter of each microscope field ( n = 4 biological replicates). Scale bar, 10 µm. I Confocal microscopy of Gal-3 (3.3 µM, containing 0.8 µM GFP-Gal-3 and 2.5 µM Gal-3) induced condensates in CHO-K1 cells expressing the mCherry-integrin α5, with or without 1, 6-hexanediol (1, 6-HD) (1.5%, 2 min). An enlarged view of the boxed region was shown on the right, with cross-sectional fluorescence intensity profiles along the white dotted line in histograms demonstrating the correlation between the two signals. Quantifications of the size and the number of the condensates per cell were shown (upper, n = 5, 5, 5 fields; lower, n = 5, 5, 4 fields, total number of cells were 60-70 in each group). Scale bar, 20 μm. J FRAP measurements. The co-localized Gal-3/integrin α5 condensates in living cells were randomly selected for bleaching (upper panel). Enlarged views of the boxed region were shown. Representative confocal microscopy images (middle panel) and normalized fluorescence intensity (lower panel) after bleaching ( n = 5 independent measurements). Scale bar, 10 μm. K Immunoblot analysis and quantifications of the phosphorylation of FAK (p-FAK, Y397) in HMEC-1 cells treated with Gal-3 (0.33 µM, 15 min) together with PBS, lactose (4 mM), or 1, 6-HD (1.5%, 2 min) in the indicated group ( n = 6, 5, 6, 6 biological replicates). L Confocal microscopy of Gal-3 (3.3 µM, containing 0.8 µM GFP-Gal-3 and 2.5 µM Gal-3) induced condensates in CHO-K1 cells co-expressing mCherry-integrin α5 and mTagBFP2-CD146 treated with lactose (10 mM), sucrose (10 mM) or 1, 6-HD (1.5%, 2 min). Enlarged views of the boxed region were shown with corresponding cross-sectional fluorescence intensity profiles along the white dotted line in histograms demonstrating the correlation between the three signals. Quantifications of the size and number of the condensates per cell were shown ( n = 5 fields, total number of cells was 40–80 in each group). Scale bar, 20 μm. M Recombinant Gal-3-induced tube formation in HMEC-1 cells treated with siRNA targeting integrin β1 or CD146 ( n = 3 biological replicates). All statistical data are presented as means ± SEM. P values were determined by unpaired two-tailed Student’s t test ( A , H , M ), one-way ANOVA with Fisher’s LSD post hoc test ( C , E , F , K ), two-sided Mann-Whitney U test ( I , L ) or Kruskal–Wallis test with Dunn’s post hoc test ( B , D ). Error bars mean ± SEM of each group. * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source Data file. Exact p values are provided in the Source Data file.

Article Snippet: After blocking with 5% non-fat dry milk in PBST (PBS containing 0.05% Tween-20), blots were incubated overnight at 4°C with the following primary antibodies: CD31 (1:1000, ab281583, Abcam, USA), β-Actin (1:1000, 612657, BD Biosciences, USA), Gal-3 (1:1000, ab76245, Abcam, USA), collagen I (1:1000, GB114197 , Servicebio, China), collagen III (1:1000, EPR17673 , Abcam, USA), AKT (1:1000, 9272 s, cell signaling technology, USA), p-AKT (Ser473) (1:1000, 9271 s, cell signaling technology, USA), FoxO1 (1:1000, 18592-1-AP, Proteintech, China), integrin β1 (1:1000, 610467, BD Biosciences, USA), integrin α3 (1:1000, 66070, Proteintech, China), integrin α5 (1:1000, sc-10729, Santa Cruz, USA), integrin α6 (1:1000, ab181551, Abcam, USA), p-integrin β1 (Y783) (1:1000, ab62337, Abcam, USA), VEGFR2 (1:1000, 2479S, Cell signaling, USA), p-VEGFR2 (Tyr1175) (1:1000, 2478S, Cell signaling technology, USA), EGFR (1:1000, 4267S, Cell signaling, USA), p-EGFR (Tyr1068) (1:1000, 3777S, Cell signaling technology, USA), integrin αv(1:1000, 27096-1-AP, Proteintech, China), FAK (1:1000, 12636-1-AP, Proteintech China), p-FAK (Y397) (1:1000, 611806, BD Biosciences, USA), CD146 (1:1000, 611208, BD Biosciences, USA), GST (1:1000, KM8005, Sungene Biotech, China), His-tag (1:1000, RM1001, Beijing Ray Antibody Biotech, China), α-Tubulin (1:1000, F0063, Selleck, USA).

Techniques: Confocal Microscopy, Recombinant, Membrane, Fluorescence, Microscopy, Spectrophotometry, Expressing, Western Blot, Phospho-proteomics, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-3-integrin α5β1 phase separation disrupted by advanced glycation end-products impairs diabetic wound healing in rodents

doi: 10.1038/s41467-025-62320-w

Figure Lengend Snippet: A Tube formation induced by recombinant Gal-3 in HUVECs with diabetic or non-diabetic patient serum (7.5%, v/v) (Non-DM, n = 5; DM, n = 6 biological replicates). B Recombinant Gal-3-induced tube formation in HUVECs treated with different concentrations of BSA-conjugated AGEs (0, 1, 10, 100 µg/mL) was normalized to the group treated with the corresponding concentration of BSA alone ( n = 3 biological replicates). C Immunoblot analysis of phosphorylated-integrin β1 (p-integrin β1) in HMEC-1 cells with Gal-3 (0.33 µM) in the presence or absence of BSA (1.98 µM) or AGEs (1.98 µM). Quantifications were shown below the blots. D Pull-down assays. HMEC-1 cell lysates (100 μg) were pulled down with His-Gal-3 in the presence or absence of BSA or AGEs, the molar ratio of Gal-3 with BSA or AGEs was 1:6. Immunoblot analysis and quantifications of integrin β1 were shown. E Chemical shift changes (Δδ) from these HSQC spectra of Gal-3 and integrin β1. 1 H- 15 N HSQC spectral expansions for 15 N-enriched Gal-3 (20 μM) in the presence of integrin β1 (0.4 μM), plus AGEs (0.4 μM, lower panel). F Bio-Layer interferometry (BLI) analysis of Gal-3-integrin β1 affinity. His-integrin β1 interacted with Gal-3 (200, 400, 600, 800, 1000, 1200, 1400 nM) (left) or Gal-3 (1.4 µM), respectively, plus different concentrations of AGEs (0, 11.2, 22.4, 44.8 µM) (right) at 25 °C. G Representative fluorescence images of condensates formed with GFP-Gal-3 (40 µM) plus BSA or AGEs (240 µM), GFP-Gal-3 (40 µM) + integrin β1 (400 nM) mixture plus BSA or AGEs (240 µM) in PBS (pH 7.4). Quantifications of condensates’ number and diameter in each microscope field were shown ( n = 4 biological replicates). Scale bar, 10 µm. H Particle size of condensates formed by Gal-3 (40 µM) and integrin β1 (400 nM) plus BSA (240 µM) or AGEs (240 µM) in PBS (pH 7.4) ( n = 3 biological replicates). I Confocal microscopy of GFP-Gal-3 (3.3 µM) induced condensates in CHO-K1 cells expressing mCherry-integrin α5 with treatment of BSA (19.8 µM) or AGEs (19.8 µM). The cell indicated by the white arrow was enlarged. Quantifications of the size and the number of the condensates per cell were shown (upper, left to right, n = 5, 6, 6, 5 fields; lower, n = 5 fields; total number of cells were 50–80 in each group). Scale bar, 20 μm. J , K STZ-induced diabetic rats were treated with hydrogels embedded AGEs inhibitor (DM + AGEi) or vehicle (DM + Veh) after wounding. Normal rats treated with blank hydrogels (Normal + Veh) served as the negative control. J Representative images of wounds and percentage of wound closure ( n = 6). K Immunohistochemical staining of CD31(black arrows) in healed wounds. Quantifications of microvessel count per field were shown on the right (left to right, n = 6, 7, 5). Scale bar, 50 µm. All statistical data are presented as means ± SEM. P values were determined by unpaired two-tailed Student’s t test ( A , G , H , J , K ), one-way ANOVA with two-sided Fisher’s LSD post hoc test ( B ) or two-sided Mann–Whitney U test ( I ). Error bars represent the mean ± SEM of each group. * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source Data file. Exact p values are provided in the Source Data file.

Article Snippet: After blocking with 5% non-fat dry milk in PBST (PBS containing 0.05% Tween-20), blots were incubated overnight at 4°C with the following primary antibodies: CD31 (1:1000, ab281583, Abcam, USA), β-Actin (1:1000, 612657, BD Biosciences, USA), Gal-3 (1:1000, ab76245, Abcam, USA), collagen I (1:1000, GB114197 , Servicebio, China), collagen III (1:1000, EPR17673 , Abcam, USA), AKT (1:1000, 9272 s, cell signaling technology, USA), p-AKT (Ser473) (1:1000, 9271 s, cell signaling technology, USA), FoxO1 (1:1000, 18592-1-AP, Proteintech, China), integrin β1 (1:1000, 610467, BD Biosciences, USA), integrin α3 (1:1000, 66070, Proteintech, China), integrin α5 (1:1000, sc-10729, Santa Cruz, USA), integrin α6 (1:1000, ab181551, Abcam, USA), p-integrin β1 (Y783) (1:1000, ab62337, Abcam, USA), VEGFR2 (1:1000, 2479S, Cell signaling, USA), p-VEGFR2 (Tyr1175) (1:1000, 2478S, Cell signaling technology, USA), EGFR (1:1000, 4267S, Cell signaling, USA), p-EGFR (Tyr1068) (1:1000, 3777S, Cell signaling technology, USA), integrin αv(1:1000, 27096-1-AP, Proteintech, China), FAK (1:1000, 12636-1-AP, Proteintech China), p-FAK (Y397) (1:1000, 611806, BD Biosciences, USA), CD146 (1:1000, 611208, BD Biosciences, USA), GST (1:1000, KM8005, Sungene Biotech, China), His-tag (1:1000, RM1001, Beijing Ray Antibody Biotech, China), α-Tubulin (1:1000, F0063, Selleck, USA).

Techniques: Recombinant, Concentration Assay, Western Blot, Fluorescence, Microscopy, Confocal Microscopy, Expressing, Negative Control, Immunohistochemical staining, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Galectin-3-integrin α5β1 phase separation disrupted by advanced glycation end-products impairs diabetic wound healing in rodents

doi: 10.1038/s41467-025-62320-w

Figure Lengend Snippet: In circulation, Gal-3 directly interacts with integrin α5β1 via glycans in vascular endothelial cells, forming a liquid-liquid phase separation, activating downstream FAK, ultimately promoting angiogenesis and skin wound healing. In diabetic states, accumulated AGEs bind to Gal-3, blocking the activation of the integrin α5β1-FAK axis, resulting in reduced angiogenesis and delayed skin wound healing. This figure was created in BioRender. Chen, S. (2025) https://BioRender.com/4tkiilw .

Article Snippet: After blocking with 5% non-fat dry milk in PBST (PBS containing 0.05% Tween-20), blots were incubated overnight at 4°C with the following primary antibodies: CD31 (1:1000, ab281583, Abcam, USA), β-Actin (1:1000, 612657, BD Biosciences, USA), Gal-3 (1:1000, ab76245, Abcam, USA), collagen I (1:1000, GB114197 , Servicebio, China), collagen III (1:1000, EPR17673 , Abcam, USA), AKT (1:1000, 9272 s, cell signaling technology, USA), p-AKT (Ser473) (1:1000, 9271 s, cell signaling technology, USA), FoxO1 (1:1000, 18592-1-AP, Proteintech, China), integrin β1 (1:1000, 610467, BD Biosciences, USA), integrin α3 (1:1000, 66070, Proteintech, China), integrin α5 (1:1000, sc-10729, Santa Cruz, USA), integrin α6 (1:1000, ab181551, Abcam, USA), p-integrin β1 (Y783) (1:1000, ab62337, Abcam, USA), VEGFR2 (1:1000, 2479S, Cell signaling, USA), p-VEGFR2 (Tyr1175) (1:1000, 2478S, Cell signaling technology, USA), EGFR (1:1000, 4267S, Cell signaling, USA), p-EGFR (Tyr1068) (1:1000, 3777S, Cell signaling technology, USA), integrin αv(1:1000, 27096-1-AP, Proteintech, China), FAK (1:1000, 12636-1-AP, Proteintech China), p-FAK (Y397) (1:1000, 611806, BD Biosciences, USA), CD146 (1:1000, 611208, BD Biosciences, USA), GST (1:1000, KM8005, Sungene Biotech, China), His-tag (1:1000, RM1001, Beijing Ray Antibody Biotech, China), α-Tubulin (1:1000, F0063, Selleck, USA).

Techniques: Blocking Assay, Activation Assay

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: Proteins affected by PM2.5-treated lung cancer cells. (A) Venn diagram showing the number of proteins expressed in the control and PM2.5-treated cells. Heatmap representing the level of expression of 61 proteins associated with “cell migration” (GO:0016477), with 9 down-regulated proteins, 7 up-regulated proteins, and 45 proteins present in both groups. (B) The protein-protein interaction (PPI) network of the down- and up-regulated proteins related to the GO term “cell migration” (GO: 0016477) in response to exposure to PM2.5. The blue circle represents the down-regulated protein, and the red circles represent the up-regulated proteins involved in cell migration. PM2.5: Particulate matter 2.5; GO: gene ontology; PTPRF: receptor-type tyrosine-protein phosphatase F; FGFR1: fibroblast growth factor receptor 1; SH3KBP1: SH3 domain-containing kinase-binding protein 1; FSCN1: fascin; ARF4: ADP-ribosylation factor 4; TYRO3: tyrosine-protein kinase receptor TYRO3.

Article Snippet: The primary antibodies against FGFR1 (#9740), integrin αV (#4711), integrin β1 (#4706), Akt (#9272), p-Akt (#4060), and β-actin (#4970), and the secondary antibody anti-rabbit IgG (#7074) were all acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Control, Expressing, Migration, Binding Assay

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: Effect of PM2.5 on proteins related to cell migration through the Rap1 signaling pathway obtained from the KEGG pathway database. Several downstream effectors are modulated by FGFR1 through integrin signaling. The red box represents proteins affected by PM2.5 treatment. PM2.5: Particulate matter 2.5; Rap1: Ras-associated protein-1; FGFR1: fibroblast growth factor receptor 1 (as members of the receptor tyrosine kinase (RTK) family); KEGG: Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: The primary antibodies against FGFR1 (#9740), integrin αV (#4711), integrin β1 (#4706), Akt (#9272), p-Akt (#4060), and β-actin (#4970), and the secondary antibody anti-rabbit IgG (#7074) were all acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Migration

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: The FGFR1-dependent pathway is a target of PM2.5 in non-small cell lung cancer H460 and A549 cells. (A-B) H460 cells were treated with PM2.5 of 0-200 μg/ml and (C-D) A549 cells were treated with PM2.5 of 0-100 μg/ml for 24 h. Then, the expression level of FGFR1 was determined by western blot analysis. β-actin was used as a loading control. The band intensities of the PM2.5 treatment groups were quantified by densitometry using ImageJ, and the results are presented as a relative protein level. (E-F) H460 cells and (G-H) A549 cells were treated with PM2.5 of 0-200 μg/ml for 24 h. The expression of FGFR1 was determined by immunofluorescence. The fluorescence intensity was analyzed by ImageJ, and the results are presented as relative protein intensity. Data are presented as mean±standard deviation (n=3). Multiple comparisons were conducted using one-way analysis of variance (ANOVA) followed by Turkey’s post hoc test and the statistical difference between the two groups was compared using t-test.*p<0.05 and **p<0.01 versus untreated control cells. PM2.5: Particulate matter 2.5.

Article Snippet: The primary antibodies against FGFR1 (#9740), integrin αV (#4711), integrin β1 (#4706), Akt (#9272), p-Akt (#4060), and β-actin (#4970), and the secondary antibody anti-rabbit IgG (#7074) were all acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Fluorescence, Standard Deviation

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: Schematic diagram that summarizes the underlying mechanism of PM2.5 that promotes lung cancer metastasis through FGFR1/integrin/Akt signaling. NSCLC: non-small cell lung cancer. FGFR1: Fibroblast growth factor receptor 1; PI3K: phosphatidylinositol 3-kinases; Akt: protein kinase B (PKB); FAK: focal adhesion kinase. Figure created with BioRender.com.

Article Snippet: The primary antibodies against FGFR1 (#9740), integrin αV (#4711), integrin β1 (#4706), Akt (#9272), p-Akt (#4060), and β-actin (#4970), and the secondary antibody anti-rabbit IgG (#7074) were all acquired from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: The FGFR1-dependent pathway is a target of PM2.5 in non-small cell lung cancer H460 and A549 cells. (A-B) H460 cells were treated with PM2.5 of 0-200 μg/ml and (C-D) A549 cells were treated with PM2.5 of 0-100 μg/ml for 24 h. Then, the expression level of FGFR1 was determined by western blot analysis. β-actin was used as a loading control. The band intensities of the PM2.5 treatment groups were quantified by densitometry using ImageJ, and the results are presented as a relative protein level. (E-F) H460 cells and (G-H) A549 cells were treated with PM2.5 of 0-200 μg/ml for 24 h. The expression of FGFR1 was determined by immunofluorescence. The fluorescence intensity was analyzed by ImageJ, and the results are presented as relative protein intensity. Data are presented as mean±standard deviation (n=3). Multiple comparisons were conducted using one-way analysis of variance (ANOVA) followed by Turkey’s post hoc test and the statistical difference between the two groups was compared using t-test.*p<0.05 and **p<0.01 versus untreated control cells. PM2.5: Particulate matter 2.5.

Article Snippet: The membranes were blocked with 5% skimmed milk in TBST (25 mm Tris-HCl, pH 7.4, 125 mm NaCl, and 0.05% Tween 20) for 1 h. They were then incubated with the specific primary antibodies against FGFR1, integrin αV, integrin β1, Akt, p-Akt and β-actin (Cell Signaling, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Fluorescence, Standard Deviation

Journal: Cancer Genomics & Proteomics

Article Title: Particulate Matter 2.5 Induces FGFR1-mediated Integrin Switch to Promote Non-small Cell Lung Cancer Metastasis

doi: 10.21873/cgp.20527

Figure Lengend Snippet: PM2.5 induces the expression of proteins involved cell migration in non-small cell lung cancer H460 and A549 cells. (A-B) H460 cells were treated with PM2.5 of 0-200 μg/ml and (C-D) A549 cells were treated with PM2.5 of 0-100 μg/ml for 24 h. Then, the expression level of integrin αV, β1, Akt, and p-Akt determined by western blot analysis. β-actin was used as a loading control. The band intensities of the PM2.5 treatment groups were quantified by densitometry using ImageJ, and the results are presented as relative protein level. Data are presented as mean ± standard deviation (n=3). Multiple comparisons were conducted using one-way analysis of variance (ANOVA) followed by Turkey’s post hoc test.*p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001 versus untreated control cells. PM2.5: Particulate matter 2.5.

Article Snippet: The membranes were blocked with 5% skimmed milk in TBST (25 mm Tris-HCl, pH 7.4, 125 mm NaCl, and 0.05% Tween 20) for 1 h. They were then incubated with the specific primary antibodies against FGFR1, integrin αV, integrin β1, Akt, p-Akt and β-actin (Cell Signaling, Danvers, MA, USA).

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