Journal: Oncogene

Article Title: A role for collagen XXIII in cancer cell adhesion, anchorage-independence, and metastasis

doi: 10.1038/onc.2011.406

Figure Lengend Snippet: Whole cell lysates were collected from H460 clonal cell lines and probed for various proteins involved in cell adhesion. In the knockdown cell lines, a reduction in OB-cadherin, gamma-, beta-, and alpha-catenin, vimentin and galectin-3 was observed. Analysis of the canonical cadherins revealed no changes in P-cadherin. E- and N-cadherin are not expressed in these cells. Actin was used as a loading control.

Article Snippet: The antibodies used were mouse monoclonal antibodies to collagen XXIII, galectin-3 (R&D Systems), integrins α2, α3, α5, β3, β-catenin, p120-catenin, E, N, and P-Cadherin, FAK, pFAK397, (BD Biosciences), actin (Chemicon), vimentin (DAKO) and rabbit antibodies to ILK1, α-catenin, γ-catenin, OB-Cadherin, AKT, Src, pSrcY416, and integrins α4, αv, α6, β1, β4, β5 (Cell Signal).

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Deficiency of Myeloid Pfkfb3 Protects Mice From Lung Edema and Cardiac Dysfunction in LPS-Induced Endotoxemia

doi: 10.3389/fcvm.2021.745810

Figure Lengend Snippet: Myeloid-specific Pfkfb3 deficiency attenuates LPS-induced inflammatory responses. (A) Representative images (left) and quantification (right) for immunohistochemical staining of the neutrophil marker Ly6G in lung sections of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 5). (B) Representative images (left) and quantification (right) for immunohistochemical staining of the macrophage marker Mac2 in lung sections of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 5). (C–E) qPCR analysis of the mRNA levels of Il1b (C) , Il6 (D) and Nos2 (E) in the lung of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 6). (F,G) ELISA analysis of Il1b (F) and Il6 (G) in serum of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 4). (H) NO levels in serum of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 3–4). All data are represented as mean ± SEM, ** P < 0.01 and *** P < 0.001 for Pfkfb3 WT vs. Pfkfb3 ΔMϕ (unpaired two-tailed Student's t test).

Article Snippet: After antigen retrieval with Antigen Unmasking Solution (H-3301, Vector Laboratories, Burlingame, CA, USA) at 98°C for 10 min, sections were blocked with avidin solution with 10% normal rabbit serum for 1 h at room temperature, and incubated in biotin blocking solution with primary antibodies against Mac2 (3 μg/mL, CL8942F, Cedarlane, Burlington, NC, USA), or Ly6G (3 μg/mL, 551459, BD biosciences, San Jose, CA, USA) at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Marker, Injection, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Proteomic Profiling of Two Distinct Populations of Extracellular Vesicles Isolated from Human Seminal Plasma

doi: 10.3390/ijms21217957

Figure Lengend Snippet: Isolation and characterization of LEV and SEV. Total EV in seminal plasma of vasectomized men were collected by UC at the interface of a sucrose block gradient. LEV and SEV were separated by their distinct velocities during upward displacement into a continuous sucrose density gradient. ( A ) Gradient fractions were analyzed by SDS-PAGE followed by Sypro ruby staining for total protein. ( B ) Gradient fractions were analyzed by immunoblotting for the presence of EV associated proteins, including CD9, CD81, PSCA, Galectin-3, CD63, HSP70, Annexin A1, and GLIPR2. Molecular weight markers are indicated on the left in kDa. ( C ) Particles in the LEV and SEV containing fractions were analyzed by TEM. Scale bar, 500 nm. Arrows exemplify incidental SEV in the LEV isolate. ( D ) Size distribution of the particles in LEV and SEV isolates as determined by NTA.

Article Snippet: The primary antibodies used were mouse anti human CD9 (HI9a, 312102, Biolegend, San Diego, California, USA, 1:2000); mouse anti human CD81 (B11, sc-166029, Santa Cruz Biotechnology, Dallas, Texas, USA, 1:500); mouse anti human HSP70 (N27F3-4, ADI-SPA-820-D, Enzo, Bruxelles, Belgium, 1:1000); mouse anti human Flotillin-1 (clone 18, 610821, BD Biosciences, San Jose, California, USA, 1:1000); mouse anti-human PSCA (clone 7F5, sc-80654, Santa Cruz Biotechnology, Dallas, Texas, USA, 1:1000); mouse anti human Annexin A1 (clone 29, 610066, BD Biosciences, San Jose, California, USA, 1:1000); mouse anti human CD47 (B6H12, sc-12730, Santa Cruz Biotechnology, Dallas, Texas, USA, 1:500); rat anti human galectin-3 (M3/38, CL8942B, CEDARLANE, Burlington, Vermont, USA, 1:500) and rabbit anti human GLIPR2/GAPR-1 (Eberle et al., 2002, 1:5000, kindly provided by J.B. Helms).

Techniques: Isolation, Blocking Assay, SDS Page, Staining, Western Blot, Molecular Weight