Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: MMP-12 (Human MMP-12 ELISA, Cat # EH327RB, Invitrogen), ADAM-9 (Human ADAM-9 DuoSet ELISA, Cat # DY939, R&D Systems), TIMP-1 (Human TIMP-1 DuoSet ELISA, Cat # DY970, R&D Systems), TIMP-2 (Human TIMP-2 DuoSet ELISA, Cat # DY971, R&D Systems), MMP-9/TIMP-1 complex (Human MMP-9/TIMP-1 DuoSet ELISA, Cat # DY1449, R&D Systems), Cystatin C (Human Cystatin C DuoSet ELISA, Cat # DY1196, R&D Systems), Cathepsin D (Human Cathepsin D DuoSet ELISA, Cat # DY1014, R&D Systems), Serpin E1 (Human Serpin E1 DuoSet ELISA, Cat # DY1786, R&D Systems), Tumor Necrosis Factor alpha (TNF-α) (Human TNF-alpha DuoSet ELISA, Cat # DY210, R&D Systems), IL-1β (Human IL-1 beta DuoSet ELISA, Cat # DY201, R&D Systems), IL-6 (Human IL-6 DuoSet ELISA, Cat # DY206, R&D Systems) and CCL-18 (Human CCL18/PARC DuoSet ELISA, Cat # DY394, R&D Systems) concentrations were measured by sandwich ELISA according to the manufacturer’s protocols.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: Anti-Human ADAM-9 , R&D systems , Cat # DY939.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: After protein transfer, membranes were blocked for 5 min in EveryBlot Blocking buffer (Cat # 12010020, Biorad) and incubated with anti-MMP-9 (Cat # AB911, R&D systems), anti-MMP-12 (Cat # AF917, R&D systems), anti-ADAM-9 (Cat # DY939, Detection antibody of Human DuoSet ELISA, R&D systems), anti-SP1 (Cat # 5931, Cell Signaling), anti-β-actin (Cat # 20536-1, Proteintech), anti-H3 (Cat # 9715, Cell Signaling), anti-CD44 (Cat # 3570, Cell Signaling), anti-HSP90 (Cat # MAB3286, R&D systems), anti-TIMP-1 (Cat # 8946S, Cell Signaling) or anti-A2M (Cat # AF1938, R&D Systems) antibodies diluted in 10% EveryBlot Blocking buffer in TBS-T (1× TBS containing 0.1% Tween 20) overnight at 4°C.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: eLife

Article Title: Proteomic landscape of tunneling nanotubes reveals CD9 and CD81 tetraspanins as key regulators

doi: 10.7554/eLife.99172

Figure Lengend Snippet: ( A ) Venn diagram showing common and exclusive proteins between Integrin adhesion complexes (blue circle) and tunneling nanotubes (TNTs) (yellow circle). The percentages refer to total proteins. ( B ) Venn diagram showing common and exclusive proteins between consensus adhesome (blue circle) and TNTs (yellow circle). ( C ) Representative immunofluorescence pictures showing expression of Integrin b1 and CD151 in TNTs of U2OS and SH-SY5Y as indicated on the left. Each picture is one upper slice of the stack, TNTs are further characterized by actin presence (actin chromobody-GFP, first lane) or wheat germ agglutinin (WGA) labeling. The yellow arrowheads point to TNTs, scale bars are 10 μm. ( D ) Representative immunofluorescence pictures showing labeling of Vinculin and Paxillin in U2OS cells cultured in complete medium or serum-free medium for 24 hr before fixation, as indicated on the left. For each labeling, the bottom slice of the stack is shown on the bottom row (z number), upper slice shows a TNT, pointed with the yellow arrowhead. Red is phalloidin staining, blue is DAPI in merge pictures; scale bars are 10 μm. ( E ) Representative immunofluorescence pictures showing labeling of Vinculin and Paxillin in SY-SY5Y cells, as in D. ( F ) Representative immunofluorescence pictures showing labeling of GM130 in U2OS cells (left, complete or serum-free medium), and SH-SY5Y cells. The yellow arrowheads point to TNTs, scale bars are 10 μm. ( G ) Expression of TM proteins in U2OS whole cell extracts (WCE), which are not in TNTome. WB from wild-type (WT) or GFP-CD9 expressing cells, incubated with the following antibodies as indicated on the left: Integrin b4 (Int b4), a4 (Int a4), EGFR and Connexin 43 (Cx43). Annexin A2 (ANXA2) is used as a loading control. WCE from all cell lines have been tested three times, two WCE are shown. ( H ) Comparative expression of proteins in WCE and TNTs from various U2OS cell lines. Left, CD9, GFP-CD9, and ADAM10 are compared in WT and GFP-CD9 expressing U2OS cells (using non-reducing gels). Right, Int b1 and ANXA2 are compared in H2B-GFP and Actin chromobodies-expressing cells (gels in reducing conditions). Figure 2—figure supplement 1—source data 1. Uncropped and labeled western blots (WBs) for . Figure 2—figure supplement 1—source data 2. Raw unedited western blots (WBs) for . Figure 2—figure supplement 1—source data 3. Uncropped and labeled western blots (WBs) for . Figure 2—figure supplement 1—source data 4. Raw unedited western blots (WBs) for .

Article Snippet: Antibody , Mouse monoclonal anti-ADAM10 11G2 , , Diaclone: #857.800.000 , WB (1/1000).

Techniques: Immunofluorescence, Expressing, Labeling, Cell Culture, Staining, Incubation, Control, Western Blot

Journal: eLife

Article Title: Proteomic landscape of tunneling nanotubes reveals CD9 and CD81 tetraspanins as key regulators

doi: 10.7554/eLife.99172

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal anti-ADAM10 11G2 , , Diaclone: #857.800.000 , WB (1/1000).

Techniques: Transfection, Construct, Expressing, Plasmid Preparation, Marker, Sequencing, Purification, Transduction, Control, Software, Staining