anti sdc1 antibody  (Human Protein Atlas)

Journal: Research

Article Title: Mapping Immune-Inflammatory Niches on Zirconia Bone Implants: Single-Cell Transcriptomic Profiling

doi: 10.34133/research.1162

Figure Lengend Snippet: Intercellular cross talk and functional enrichment in the bone-marrow microenvironment following titanium (Ti) implantation. (A and B) Cells extracted from the Ti implant (A) and their proportions compared with those in the sham group (B). (C) Fuzzy C -means clustering of the differentially expressed genes (DEGs) and their functional annotation via Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses. BP, Biological Process; CC, Cellular Component; MF, Molecular Function. (D and E) Outgoing–incoming cross talk analysis among the bone-marrow cells and their subpopulations. (F) Outgoing–incoming signaling patterns and associated interaction strengths. (G) Analysis of collagen-related signaling networks and intercellular interaction strengths. (H) Interaction-strength analysis of candidate ligand–receptor pairs in bone-marrow fibroblasts. (I) KEGG and GO (BP, CC, and MF) enrichment analyses of the DEGs in bone-marrow fibroblasts. (J and K) Multiplex fluorescence colocalization and costaining analysis of candidate ligand–receptor pairs. Scale bars: 100 and 20 μm. COL1A1, collagen type I alpha 1 chain; SDC1, syndecan 1; ATP, adenosine triphosphate; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: The following antibodies were applied sequentially: CD44 (Bioss, bsm-54767R, 1:300, China), CD68 (Proteintech, 66231-2-Ig, 1:500, China), CD206 (Bioss, bsm-55604R, 1:300, China), NOS2 (Bioss, bs-0162R, 1:300, China), COL6A2 (Bioss, bs-13963R, 1:300, China), S100A4 (Bioss, bs-3759R, 1:300, China), COL1A1 (Bioss, bs-10423R, 1:300, China), and SDC1 (Bioss, bs-1309R, 1:300, China), followed by nuclear counterstaining with 4′,6-diamidino-2-phenylindole.

Techniques: Functional Assay, Multiplex Assay, Fluorescence

Journal: Research

Article Title: Mapping Immune-Inflammatory Niches on Zirconia Bone Implants: Single-Cell Transcriptomic Profiling

doi: 10.34133/research.1162

Figure Lengend Snippet: Validation of the specific ligand–receptor pairs between the implants and bone-marrow cells. (A) Schematic diagram of the rat model of intramedullary femoral implantation and bulk RNA sequencing workflow. (B) Violin plots showing the expression levels of Col6a2 , Cd44 , Col1a1 , and Sdc1 . (C and D) Hierarchical clustering heatmaps of pairwise DEGs among the sham, Ti, and ZrO 2 groups. (E to G) Identification of the gene modules associated with Ti and ZrO 2 via weighted gene coexpression network analysis. (H) Overlapping genes between module-related genes and DEGs in each group. (I) Contributions of Ti and ZrO 2 -associated feature genes to predictive outcomes, assessed using least absolute shrinkage and selection operator (LASSO) regression analysis. (J and K) Temporal expression patterns and correlations of Col6a2 , Cd44 , Col1a1 , and Sdc1 . Statistical significance was assessed using the t test.

Article Snippet: The following antibodies were applied sequentially: CD44 (Bioss, bsm-54767R, 1:300, China), CD68 (Proteintech, 66231-2-Ig, 1:500, China), CD206 (Bioss, bsm-55604R, 1:300, China), NOS2 (Bioss, bs-0162R, 1:300, China), COL6A2 (Bioss, bs-13963R, 1:300, China), S100A4 (Bioss, bs-3759R, 1:300, China), COL1A1 (Bioss, bs-10423R, 1:300, China), and SDC1 (Bioss, bs-1309R, 1:300, China), followed by nuclear counterstaining with 4′,6-diamidino-2-phenylindole.

Techniques: Biomarker Discovery, RNA Sequencing, Expressing, Selection

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: Antibodies against LC3 (14600-1-AP), LAMP2 (66301-1-Ig), p62 (18420-1-AP), Syntenin (22399-1-AP), CD63 (25682-1-AP), TSG101 (28283-1-AP), CD9 (20597-1-AP), α-tubulin (11,224–1-AP), PSMA6 (67695-1-Ig), PSMED2 (14748-1-AP), USP14 (67746-1-Ig), and SDC1 (10593-1-AP) for SDC1-FL detection were obtained from Proteintech. β-catenin (ab16051) and PSMD14 (ab109123) antibodies were obtained from Abcam.

Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: SDC4-CTF is further cleaved by γ-secretase to release the cytoplasmic fragment. A , confocal microscopy of HCT116 cells transfected with SDC4-GFP or SDC4-CTF-GFP. B , schematic of SDC4-GFP deletion mutants, SNP for Signal peptide, ED for ectodomain, TM for TM motif, C1 and C2 for the constant regions, and V for variable region. C , Western blot of HCT116 cells transfected with SDC1-GFP, SDC4-GFP, or SDC4-CTF-GFP, with/without 5 μM MG132 (4 h). D , sequence alignment of Syndecan family TM C-terminal fragments. The alignment was performed using Clustal W, and analysis was done with ESPript 3.0. E , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with presenilin-1 knockdown (siRNA, 72 h). F , Western blot of HCT116 and SW480 cells treated with 10 μM DAPT, 10 μM MG132, or 2.5 μM GM6001 (12 h). G , schematic of SDC4-CTF degradation via lysosome or proteasome pathways. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment.

Article Snippet: Antibodies against LC3 (14600-1-AP), LAMP2 (66301-1-Ig), p62 (18420-1-AP), Syntenin (22399-1-AP), CD63 (25682-1-AP), TSG101 (28283-1-AP), CD9 (20597-1-AP), α-tubulin (11,224–1-AP), PSMA6 (67695-1-Ig), PSMED2 (14748-1-AP), USP14 (67746-1-Ig), and SDC1 (10593-1-AP) for SDC1-FL detection were obtained from Proteintech. β-catenin (ab16051) and PSMD14 (ab109123) antibodies were obtained from Abcam.

Techniques: Confocal Microscopy, Transfection, Western Blot, Sequencing, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: Syntenin stabilizes SDC4-CTF against endocytic-lysosomal degradation. A , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 25 μM LY294002, 250 nM Rapamycin, or 250 nM Bafilomycin A1. B , Western blot of HCT116 cells starved in EBSS for indicated durations. C and D , Western blot and quantification of SDC1-CTF and SDC4-CTF in HCT116 cells overexpressing Flag-tagged Syntenin (n = 3). E , schematic and sequences of SDC4 WT and SDC4 ΔC2 , with confocal images of indicated plasmids in 293T cells. F , quantification of fluorescence co-localization from ( E ) (n = 6). G and H , Western blot and quantification of SDC4-CTF and Syntenin in 293T cells co-transfected with SDC4 and GFP-tagged Syntenin (n = 3). I , Western blot of SDC1-CTF, SDC4-CTF, and Syntenin in multiple cancer cell lines. J , correlation analysis between SDC4-CTF and Syntenin expression from ( I ). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: Antibodies against LC3 (14600-1-AP), LAMP2 (66301-1-Ig), p62 (18420-1-AP), Syntenin (22399-1-AP), CD63 (25682-1-AP), TSG101 (28283-1-AP), CD9 (20597-1-AP), α-tubulin (11,224–1-AP), PSMA6 (67695-1-Ig), PSMED2 (14748-1-AP), USP14 (67746-1-Ig), and SDC1 (10593-1-AP) for SDC1-FL detection were obtained from Proteintech. β-catenin (ab16051) and PSMD14 (ab109123) antibodies were obtained from Abcam.

Techniques: Western Blot, Fluorescence, Transfection, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO binds to SDC4 and disrupts SDC4-Syntenin interaction. A , chemical structure of AGO. B , microscale thermophoresis (MST) assays of AGO with GFP-SDC4 or GFP control in 293T lysate (n = 3). C , MST assays of AGO with recombinant glutathione S-transferase (GST)-SDC4 (n = 3). D and E , MST assays of AGO with GFP-tagged SDC4 or SDC1 and their TM deletion mutants (n = 3 each). F and G , Western blot and quantification of AGO drug affinity responsive target stability experiments (n = 3). H and I , immunoprecipitation (IP) and quantification of SDC4 binding to Syntenin in SW480 cells treated with 20 μM AGO (24 h; n = 3). J , IP of Flag-Syntenin and GFP-SDC4 in 293T cells treated with 20 μM AGO (24 h). K , GST pull-down of Syntenin from 293T lysate with/without 20 μM AGO, using GST or GST-SDC4. L and M , poximity ligation assays and quantification of SDC4-Syntenin interaction in SW480 cells treated with 20 μM AGO (24 h; n = 8). O and P , confocal images and quantification of SDC4/GFP-Syntenin co-localization in 293T cells treated with 20 μM AGO (24 h; n = 8). Q , IP of SDC4 and detection of GFP-Syntenin/endogenous Syntenin in 293T cells treated with 20 μM AGO (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). AGO, Andrographolide; GST, glutathione S-transferase; SDC4, Syndecan4; MST, microscale thermophoresis; TM, transmembrane; IP, immunoprecipitation.

Article Snippet: Antibodies against LC3 (14600-1-AP), LAMP2 (66301-1-Ig), p62 (18420-1-AP), Syntenin (22399-1-AP), CD63 (25682-1-AP), TSG101 (28283-1-AP), CD9 (20597-1-AP), α-tubulin (11,224–1-AP), PSMA6 (67695-1-Ig), PSMED2 (14748-1-AP), USP14 (67746-1-Ig), and SDC1 (10593-1-AP) for SDC1-FL detection were obtained from Proteintech. β-catenin (ab16051) and PSMD14 (ab109123) antibodies were obtained from Abcam.

Techniques: Microscale Thermophoresis, Control, Recombinant, Western Blot, Immunoprecipitation, Binding Assay, Ligation, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: Antibodies against LC3 (14600-1-AP), LAMP2 (66301-1-Ig), p62 (18420-1-AP), Syntenin (22399-1-AP), CD63 (25682-1-AP), TSG101 (28283-1-AP), CD9 (20597-1-AP), α-tubulin (11,224–1-AP), PSMA6 (67695-1-Ig), PSMED2 (14748-1-AP), USP14 (67746-1-Ig), and SDC1 (10593-1-AP) for SDC1-FL detection were obtained from Proteintech. β-catenin (ab16051) and PSMD14 (ab109123) antibodies were obtained from Abcam.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: SDC4-CTF is further cleaved by γ-secretase to release the cytoplasmic fragment. A , confocal microscopy of HCT116 cells transfected with SDC4-GFP or SDC4-CTF-GFP. B , schematic of SDC4-GFP deletion mutants, SNP for Signal peptide, ED for ectodomain, TM for TM motif, C1 and C2 for the constant regions, and V for variable region. C , Western blot of HCT116 cells transfected with SDC1-GFP, SDC4-GFP, or SDC4-CTF-GFP, with/without 5 μM MG132 (4 h). D , sequence alignment of Syndecan family TM C-terminal fragments. The alignment was performed using Clustal W, and analysis was done with ESPript 3.0. E , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with presenilin-1 knockdown (siRNA, 72 h). F , Western blot of HCT116 and SW480 cells treated with 10 μM DAPT, 10 μM MG132, or 2.5 μM GM6001 (12 h). G , schematic of SDC4-CTF degradation via lysosome or proteasome pathways. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Confocal Microscopy, Transfection, Western Blot, Sequencing, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: Syntenin stabilizes SDC4-CTF against endocytic-lysosomal degradation. A , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 25 μM LY294002, 250 nM Rapamycin, or 250 nM Bafilomycin A1. B , Western blot of HCT116 cells starved in EBSS for indicated durations. C and D , Western blot and quantification of SDC1-CTF and SDC4-CTF in HCT116 cells overexpressing Flag-tagged Syntenin (n = 3). E , schematic and sequences of SDC4 WT and SDC4 ΔC2 , with confocal images of indicated plasmids in 293T cells. F , quantification of fluorescence co-localization from ( E ) (n = 6). G and H , Western blot and quantification of SDC4-CTF and Syntenin in 293T cells co-transfected with SDC4 and GFP-tagged Syntenin (n = 3). I , Western blot of SDC1-CTF, SDC4-CTF, and Syntenin in multiple cancer cell lines. J , correlation analysis between SDC4-CTF and Syntenin expression from ( I ). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Fluorescence, Transfection, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO binds to SDC4 and disrupts SDC4-Syntenin interaction. A , chemical structure of AGO. B , microscale thermophoresis (MST) assays of AGO with GFP-SDC4 or GFP control in 293T lysate (n = 3). C , MST assays of AGO with recombinant glutathione S-transferase (GST)-SDC4 (n = 3). D and E , MST assays of AGO with GFP-tagged SDC4 or SDC1 and their TM deletion mutants (n = 3 each). F and G , Western blot and quantification of AGO drug affinity responsive target stability experiments (n = 3). H and I , immunoprecipitation (IP) and quantification of SDC4 binding to Syntenin in SW480 cells treated with 20 μM AGO (24 h; n = 3). J , IP of Flag-Syntenin and GFP-SDC4 in 293T cells treated with 20 μM AGO (24 h). K , GST pull-down of Syntenin from 293T lysate with/without 20 μM AGO, using GST or GST-SDC4. L and M , poximity ligation assays and quantification of SDC4-Syntenin interaction in SW480 cells treated with 20 μM AGO (24 h; n = 8). O and P , confocal images and quantification of SDC4/GFP-Syntenin co-localization in 293T cells treated with 20 μM AGO (24 h; n = 8). Q , IP of SDC4 and detection of GFP-Syntenin/endogenous Syntenin in 293T cells treated with 20 μM AGO (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). AGO, Andrographolide; GST, glutathione S-transferase; SDC4, Syndecan4; MST, microscale thermophoresis; TM, transmembrane; IP, immunoprecipitation.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Microscale Thermophoresis, Control, Recombinant, Western Blot, Immunoprecipitation, Binding Assay, Ligation, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: SDC4-CTF is further cleaved by γ-secretase to release the cytoplasmic fragment. A , confocal microscopy of HCT116 cells transfected with SDC4-GFP or SDC4-CTF-GFP. B , schematic of SDC4-GFP deletion mutants, SNP for Signal peptide, ED for ectodomain, TM for TM motif, C1 and C2 for the constant regions, and V for variable region. C , Western blot of HCT116 cells transfected with SDC1-GFP, SDC4-GFP, or SDC4-CTF-GFP, with/without 5 μM MG132 (4 h). D , sequence alignment of Syndecan family TM C-terminal fragments. The alignment was performed using Clustal W, and analysis was done with ESPript 3.0. E , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with presenilin-1 knockdown (siRNA, 72 h). F , Western blot of HCT116 and SW480 cells treated with 10 μM DAPT, 10 μM MG132, or 2.5 μM GM6001 (12 h). G , schematic of SDC4-CTF degradation via lysosome or proteasome pathways. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Confocal Microscopy, Transfection, Western Blot, Sequencing, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: Syntenin stabilizes SDC4-CTF against endocytic-lysosomal degradation. A , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 25 μM LY294002, 250 nM Rapamycin, or 250 nM Bafilomycin A1. B , Western blot of HCT116 cells starved in EBSS for indicated durations. C and D , Western blot and quantification of SDC1-CTF and SDC4-CTF in HCT116 cells overexpressing Flag-tagged Syntenin (n = 3). E , schematic and sequences of SDC4 WT and SDC4 ΔC2 , with confocal images of indicated plasmids in 293T cells. F , quantification of fluorescence co-localization from ( E ) (n = 6). G and H , Western blot and quantification of SDC4-CTF and Syntenin in 293T cells co-transfected with SDC4 and GFP-tagged Syntenin (n = 3). I , Western blot of SDC1-CTF, SDC4-CTF, and Syntenin in multiple cancer cell lines. J , correlation analysis between SDC4-CTF and Syntenin expression from ( I ). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Fluorescence, Transfection, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO binds to SDC4 and disrupts SDC4-Syntenin interaction. A , chemical structure of AGO. B , microscale thermophoresis (MST) assays of AGO with GFP-SDC4 or GFP control in 293T lysate (n = 3). C , MST assays of AGO with recombinant glutathione S-transferase (GST)-SDC4 (n = 3). D and E , MST assays of AGO with GFP-tagged SDC4 or SDC1 and their TM deletion mutants (n = 3 each). F and G , Western blot and quantification of AGO drug affinity responsive target stability experiments (n = 3). H and I , immunoprecipitation (IP) and quantification of SDC4 binding to Syntenin in SW480 cells treated with 20 μM AGO (24 h; n = 3). J , IP of Flag-Syntenin and GFP-SDC4 in 293T cells treated with 20 μM AGO (24 h). K , GST pull-down of Syntenin from 293T lysate with/without 20 μM AGO, using GST or GST-SDC4. L and M , poximity ligation assays and quantification of SDC4-Syntenin interaction in SW480 cells treated with 20 μM AGO (24 h; n = 8). O and P , confocal images and quantification of SDC4/GFP-Syntenin co-localization in 293T cells treated with 20 μM AGO (24 h; n = 8). Q , IP of SDC4 and detection of GFP-Syntenin/endogenous Syntenin in 293T cells treated with 20 μM AGO (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). AGO, Andrographolide; GST, glutathione S-transferase; SDC4, Syndecan4; MST, microscale thermophoresis; TM, transmembrane; IP, immunoprecipitation.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Microscale Thermophoresis, Control, Recombinant, Western Blot, Immunoprecipitation, Binding Assay, Ligation, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: SDC1 (12,922) for SDC1-CTF detection, PSMB5 (12,919), Phospho-p65 (3033), Rab5 (3547), and GAPDH (5174) antibodies were obtained from Cell Signaling Technology.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: bioRxiv

Article Title: ADAM10 tailors extracellular vesicles for content transfer rather than signaling by contact

doi: 10.64898/2026.02.12.705562

Figure Lengend Snippet: A . Left: Scheme illustrating the complexity of SDC1 structure and processing. (Upper part) SDC1 full-length (FL) core protein (black) is substituted with glycosaminoglycan chains (GAG) of the heparan (green) and chondroitin (orange) sulfate type. The trimming of GAG chains by specific enzymes is necessary for the FL protein to migrate at a discrete band in SDS-PAGE and to be detectable by immunoblotting. (Lower part) SDC1 core protein can be cleaved by proteases generating two main fragments: an N-terminal fragment comprising most of the GAG-substituted extracellular domain (ECD) and a C-terminal fragment (CTF) comprising the remainder of the ECD, the membrane-spanning and the cytoplasmic domain. Right: Western blot illustrating the signals obtained, after GAG-digestion, for FL SDC1 and SDC1 CTF in the cells and the sEV enriched fraction obtained after differential ultracentrifugation of the conditioned extracellular media. Signals were obtained with an antibody recognizing the intracellular domain of SDC1. Note that the FL form of SDC1 (SDC1-FL) abounds in cell lysates, while the CTF is less abundant. On the contrary, the SDC1-CTF is abundant and the SDC1-FL is barely detectable in sEVs. B. MCF7 cells downregulated for ADAM10 (siADAM10) or ADAM17 (siADAM17) and control cells (siCTRL) were evaluated for SDC1 FL and CTF abundance in cells and sEVs by Western blot, after GAG-digestion. Histograms represent the mean signal intensity for indicated proteins relative to the signal in control cells, ± SEM. Statistical analysis was performed using the Kruskal-Wallis one-way non-parametric ANOVA test (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, n.s. non-significant). C. MCF7 cells downregulated for ADAM10 (siADAM10) and control cells (siCTRL) were treated with heparitinase and chondroitinase (GAG digestion +) or not (GAG digestion -) to evaluate SDC substitution with GAG chains. SDCs FL and CTF in cells and sEVs were analyzed by Western blot as indicated. Single blots examining the relative abundance of FL versus CTF forms of SDC1 ( D ) or SDC4 ( E ) forms are provided. F . sEVs secreted by MCF7 cells inhibited for ADAM10 activity, (GI254023X) versus controls (DMSO) were isolated from conditioned media by differential ultracentrifugation. sEVs were analyzed by Western blot after GAG-digestion to evaluate the levels of SDC1 FL and CTF. Histograms represent the mean signal intensity for indicated proteins relative to the signal in control cells, ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, n.s. non-significant (unpaired non-parametric Mann–Whitney test). n indicates the number of independent experiments; each point represents one independent experiment.

Article Snippet: Antibodies directed against SDC1 intracellular domain (D4Y7H) was from (cell signaling #12922, dilution 1/1000), GFP (A11122) from Thermofisher (dilution 1/1000), ADAM17 (abcam, #ab39162, 1/1000 or cell signaling #3976, 1/1000), ADAM10 (abcam, #ab1997, 1/1 000), GAPDH (Proteintech # 10494-1-AP), Flotillin-1 (BD Biosciences, #X11669), Ephrin B3 (santa cruz, # sc-514139), PTK7 (Proteintech, #17799), E-cadherin (R&D Systems, #AF748), BCAM (R&D Systems, #BAF148), STAT3 124H6 (cell signaling #9139), p-STAT3 (cell signaling #9131).

Techniques: SDS Page, Western Blot, Membrane, Control, Activity Assay, Isolation, MANN-WHITNEY

Journal: bioRxiv

Article Title: ADAM10 tailors extracellular vesicles for content transfer rather than signaling by contact

doi: 10.64898/2026.02.12.705562

Figure Lengend Snippet: sEVs secreted by MCF7 ( A, D ) MDA-MB-468 ( B, E ) or SKBR3 ( C-F ) cells with knock-out of ADAM10 (A10 KO) or control cells (WT) were isolated from conditioned culture media by differential ultracentrifugation. sEVs were analyzed by Western blot after GAG-digestion to evaluate the levels of SDC1 and SDC4 FL and CTF. Histograms under A-C represent the mean signal intensity for indicated proteins relative to the signal in ADAM10-KO cells for SDC-FL and to the signal in control cells for SDC-CTF, ± SEM. The value ‘n’ indicates the number of independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, n.s. non-significant (multiple unpaired t-test). D-F. The number sEVs secreted by MCF7 ( D ), MDA-MB-468 ( E ), SKBR3 ( F ) cells knock-out for ADAM10 (ADAM10-KO) or control cells (WT) were further analyzed by microfluidic resistive pulse sensing (MRPS) - Spectradyne (nCS2). n indicates the number of independent experiments.

Article Snippet: Antibodies directed against SDC1 intracellular domain (D4Y7H) was from (cell signaling #12922, dilution 1/1000), GFP (A11122) from Thermofisher (dilution 1/1000), ADAM17 (abcam, #ab39162, 1/1000 or cell signaling #3976, 1/1000), ADAM10 (abcam, #ab1997, 1/1 000), GAPDH (Proteintech # 10494-1-AP), Flotillin-1 (BD Biosciences, #X11669), Ephrin B3 (santa cruz, # sc-514139), PTK7 (Proteintech, #17799), E-cadherin (R&D Systems, #AF748), BCAM (R&D Systems, #BAF148), STAT3 124H6 (cell signaling #9139), p-STAT3 (cell signaling #9131).

Techniques: Knock-Out, Control, Isolation, Western Blot

Journal: bioRxiv

Article Title: ADAM10 tailors extracellular vesicles for content transfer rather than signaling by contact

doi: 10.64898/2026.02.12.705562

Figure Lengend Snippet: A. sEVs secreted by SDC1-downregulated cells (siSDC1) versus controls (siCTRL) were isolated by differential ultracentrifugation. sEVs were further analyzed by Western blot, testing for several markers, as indicated. Histograms represent mean signal intensities in sEVs relative to signals obtained for controls, ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.005, **** P < 0.001, n.s. non-significant (two-way ANOVA). B. sEVs secreted by MCF7 cells downregulated for SDC1 (siSDC1), SDC4 (siSDC4) or control cells (siCTRL) were isolated by differential ultracentrifugation from conditioned media and further analyzed by Nanosight (NTA) to determine concentration and size, as indicated. Each point represents one independent experiment. C. (Upper part) Representative confocal micrographs showing the steady-state distribution of endogenous ADAM10 (red in merge) in MCF7 cells transfected with control (siCTRL) siRNA or siRNA targeting SDC1 (siSDC1). SDC1 was stained with antibody recognizing intracellular domain of SDC1 (green in merge). (Lower part) Representative confocal micrographs showing the steady-state distribution of endogenous ADAM10 (green in merge) together with CD63 (red in merge) upon SDC1 downregulation. In merge, nuclei are stained with DAPI (blue). D. sEVs secreted by cells treated with the ADAM10 inhibitor GI254023X versus DMSO controls were isolated by differential ultracentrifugation. Cell lysates from parent cells or sEVs were further analyzed by Western blot, testing for several markers, as indicated. Note that ADAM10 inhibition does not impact the loading of tetraspanins CD63, CD81 and CD9. Histograms represent mean signal intensities relative to signals obtained for controls, ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. non-significant (two-way ANOVA). The value ‘n’ indicates the number of independent experiments.

Article Snippet: Antibodies directed against SDC1 intracellular domain (D4Y7H) was from (cell signaling #12922, dilution 1/1000), GFP (A11122) from Thermofisher (dilution 1/1000), ADAM17 (abcam, #ab39162, 1/1000 or cell signaling #3976, 1/1000), ADAM10 (abcam, #ab1997, 1/1 000), GAPDH (Proteintech # 10494-1-AP), Flotillin-1 (BD Biosciences, #X11669), Ephrin B3 (santa cruz, # sc-514139), PTK7 (Proteintech, #17799), E-cadherin (R&D Systems, #AF748), BCAM (R&D Systems, #BAF148), STAT3 124H6 (cell signaling #9139), p-STAT3 (cell signaling #9131).

Techniques: Isolation, Western Blot, Control, Concentration Assay, Transfection, Staining, Inhibition

Journal: bioRxiv

Article Title: ADAM10 tailors extracellular vesicles for content transfer rather than signaling by contact

doi: 10.64898/2026.02.12.705562

Figure Lengend Snippet: A-B. Western blot of the cell lysates and sEVs from HEK293 cells treated with the ADAM10 inhibitor GI254023X or DMSO as control, in the presence or absence of serum (FCS), show that ADAM10 supports the cleavage of various receptors, as indicated by the observed increase of non-cleaved receptors in the sEV of those cells, irrespectively of serum (although the absence of serum significantly reduces sEV accumulations). These control experiments were performed to ensure the pertinence of the endosomal escape assay as developed by Hyka et al., 2025 in the context of the present study. For SDCs (A), cells were treated with heparatinase and chondroitinase (GAG digestion +) or not (GAG digestion -). Heparan sulfate proteoglycans / SDCs are detected with the mAb 3G10 recognizing GAG chain stubs left after digestion ( David et al , 1992 ). C. Left. Histogram illustrating that the uptake of HiBiT-syntenin sEVs by HEK293 LgBiT cells after 4 hours of incubation is comparable regardless of treatment with the ADAM10 inhibitor GI254023X. Right. Histogram illustrating that endosomal escape of HiBit-syntenin sEVs is abolished when sEVs originate from HEK293 treated with ADAM10 inhibitor (GI254023X). Histograms represent mean signal intensities in relative light units (RLU) after addition of the nanoluciferase substrate Furimazine. D. Illustrative western blot of size exclusion chromatography (SEC) experiments for the preparation of sEVs for functional assays. Left. Syntenin, CD9, CD81, CD63, ADAM10, SDC1 and SDC4 were used as markers to select ad-hoc fractions from the conditioned media of MCF7 cells KO for ADAM10 (sEV KO) or control cells (sEV WT) as indicated. Fractions 6-9 were pooled before incubation with HUVEC cells. Right. Syntenin, CD9, CD81, CD63, SDC1 and EGFR were used as markers to select ad-hoc fractions from the conditioned media of MDA-MB-468 WT cells as indicated. Fractions 6-8 were pooled before incubation with HUVEC cells. E. The phosphorylation of selected proteins (top nine proteins showing increased phosphorylation after ADAM10-KO EV treatment) were determined using the phospho-array (R&D systems). Signals for each phosphorylated protein are presented as a pair of duplicate spots for the same exposition of 15 minutes. Average densitometric values for the phosphorylated proteins are shown in the heatmap . F. Heatmap showing log 2-fold changes of protein phosphorylation after treatment with sEV from MDA-MB-468 ADAM10 KO cells compared to control WT cells. The log 2-fold changes of protein phosphorylation after sEV KO treatment compared to control (sEV WT) were shown in the heatmap. The level of average densitometric values (Log2) are presented as a spectrum of color where white and red colors represent the lowest and the highest values in the row according to the indicated scale. The difference (diff, shown in ochre) corresponds to the difference between KO and WT conditions of the same protein, based on their respective signal intensity difference between KO and Ctrl conditions. The experiment was performed two or three times depending on the analyzed protein.

Article Snippet: Antibodies directed against SDC1 intracellular domain (D4Y7H) was from (cell signaling #12922, dilution 1/1000), GFP (A11122) from Thermofisher (dilution 1/1000), ADAM17 (abcam, #ab39162, 1/1000 or cell signaling #3976, 1/1000), ADAM10 (abcam, #ab1997, 1/1 000), GAPDH (Proteintech # 10494-1-AP), Flotillin-1 (BD Biosciences, #X11669), Ephrin B3 (santa cruz, # sc-514139), PTK7 (Proteintech, #17799), E-cadherin (R&D Systems, #AF748), BCAM (R&D Systems, #BAF148), STAT3 124H6 (cell signaling #9139), p-STAT3 (cell signaling #9131).

Techniques: Western Blot, Control, Incubation, Size-exclusion Chromatography, Functional Assay, Phospho-proteomics