Journal: PLoS Pathogens

Article Title: Binding of Glutathione to Enterovirus Capsids Is Essential for Virion Morphogenesis

doi: 10.1371/journal.ppat.1004039

Figure Lengend Snippet: Effect of TP219-, BSO- and mock treatment on the levels of 5S protomers and 14S pentamers as determined by immunoprecipitation ( A–B ) and immunoblotting analysis ( C–E ). ( A–B ) Two consecutive fractions of a 6 to 25% gradient were pooled, immunoprecipitated using polyclonal anti-enterovirus antibodies and assayed for radioactivity. Different immunoprecipitated fractions (bars) are overlaid on TCA-precipitated fractions (lines) (derived from ) and the amount of radioactivity is expressed as % of total signal to normalize for the incorporation efficiency. Immunoprecipitated fractions of untreated cultures ( A and B ) are indicated in white; TP219- ( A ) or BSO- ( B ) treated cultures in black. Cartoon representations of the 5S and 14S assembly intermediates are indicated as well. ( C–E ) Two consecutive fractions of a 6 to 25% gradient were pooled, TCA-precipitated and subjected to Western blotting analysis using a monoclonal anti-VP1 antibody. Both the immunoblot (below) and quantification of the immunoblot (top) of mock ( C ), TP219- ( D ) and BSO- ( E ) treated cultures are depicted. Sucrose density is indicated as well and expressed as g/cm 3 .

Article Snippet: For immunoprecipitation of sucrose gradients we used pooled rabbit polyclonal anti-enterovirus antibodies (including CVB1, 2 and 3) (Accurate Chemical and Scientific Corporation).

Techniques: Immunoprecipitation, Western Blot, Radioactivity, Derivative Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Tissue inhibitor of metalloproteinases-1-induced scattered liver metastasis is mediated by host-derived urokinase-type plasminogen activator

doi: 10.1111/j.1582-4934.2009.00951.x

Figure Lengend Snippet: (A and B) Quantification of macrophages (A) and neutrophils (B) in metastasis-bearing bearing livers of META/Bom nu/nu mice (uPA +/+ and uPA −/− , respectively), transduced by either AdTIMP-1 or Addl70–3 adenoviruses, exploiting immunohistochemistry of FIRE (A) and Ly-6G (B) protein, respectively, on liver cryo-sections (5 mm). In livers with elevated TIMP-1 increased infiltration of both macrophages (A) and neutrophils (B) was detected. Loss of host uPA only diminished the number of invaded neutrophils (B), but did not impair macrophage recruitment (A). Columns : Mean number of either invaded macrophages (A) or invaded neutrophils (B) per 1.0 mm 2 liver section surface. Bars : S.E. n = 5 mice. (A) Addl70–3/uPA +/+ : 3.4 ± 0.4; Addl70–3/uPA −/− : 4.7 ± 0.3; AdTIMP-1/uPA +/+ : 6.3 ± 0.1; AdTIMP-1/uPA −/− : 5.3 ± 0.6. (B) Addl70–3/uPA +/+ : 27.8 ± 3.5; Addl70–3/uPA −/− : 26.9 ± 2.5; AdTIMP-1/uPA +/+ : 167.0 ± 2.7; AdTIMP-1/uPA −/− : 39.1 ± 7.0. (C) Representative Western blot of phosphorylated c-Met in liver protein of META/Bom nu/nu mice (uPA +/+ and uPA −/− , respectively) receiving AdTIMP-1 adenovirus. Analysis revealed similarly weak c-Met phosphorylation in livers of uPA −/− animals without depletion of neutrophils (uPA −/− /Ø) and of neutrophil depleted wild-type littermates (uPA +/+ /depleted). (D) Close-up pictures of surfaces of representative X-Gal-stained metastasis-bearing livers of uPA −/− /Ø and uPA +/+ /depleted mice, both with elevated TIMP-1 levels. Areas with one large metastasis (indigo-blue) and the immediate environment are shown. Characteristic TIMP-1-induced scattering of tumour cells (indigo-blue signal) in the livers of uPA +/+ animals (compare ) is similarly abrogated in conditions without elevated TIMP-1 levels and after depletion of neutrophils.

Article Snippet: In short, mice were inoculated with 250 μl of anti-Ly-6G rabbit antibody serum (polyclonal; Accurate Chemical & Scientific Corporation, Westbury, NY, USA) 24 hrs and 2 hrs before as well as 5 days after administration of AdTIMP-1, respectively.

Techniques: Immunohistochemistry, Western Blot, Staining