Journal:

Article Title: Anti-inflammatory effects of a new tumour necrosis factor-alpha (TNF-?) inhibitor (CNI-1493) in collagen-induced arthritis (CIA) in rats

doi: 10.1046/j.1365-2249.1999.00750.x

Figure Lengend Snippet: Immunostaining of synovial tissue in CNI-1493-treated and non-treated DA rats with CIA. Immunostaining of TNF-α (A,C) and MHC II (B,D) in knee joint tissue from rats with CIA. Synovitis, articular cartilage and bone (dark blue staining) are evident in all figures. (A,B) Stainings in consecutive sections from an animal treated daily with CNI-1493 from start of immunization (rat V in Table 3). (C,D) Consecutive sections in tissue from a placebo-treated animal (rat I in Table 3). Both animals were killed on day 21 post-immunization (p.i.), when maximal signs of inflammation occurred in the placebo-treated group of animals. A massive infiltration of MHC II+ macrophages was recorded in the synovitis of both the CNI-treated (B) and the placebo-treated (D) rat, Yet TNF-α-expressing cells were only abundant in the placebo-treated animal (C), while the number of these cells was dramatically reduced in the CNI-treated rat (A).

Article Snippet: After additional thorough washes in BSS–saponin, sections were incubated overnight at room temperature in a humidified chamber with 50 μl of cytokine-specific antigen affinity-purified antibody (either polyclonal rabbit anti-rat TNF-α (lot no. 8-14; Dr P. van der Meide, Biomedical Primate Research Centre, Rijswijk, The Netherlands), or polyclonal antigen affinity-purified goat anti-rat TNF-α (AF-510; R&D Systems, Minneapolis, MN; used at 2 μg/ml).

Techniques: Immunostaining, Staining, Expressing

Journal: Redox Biology

Article Title: A role for peroxiredoxins in H 2 O 2 - and MEKK-dependent activation of the p38 signaling pathway

doi: 10.1016/j.redox.2019.101340

Figure Lengend Snippet: Mammalian MEKK4 is redox-sensitive and interacts with cytosolic peroxiredoxin Prx2 in response to H 2 O 2 . ( A, A ′) A tagged version of Prx2 (Prx2-SBP) was expressed in HEK293T cells. Cells were treated with H 2 O 2 (100 μM) and lysed at indicated time points. Prx2-SBP was affinity-purified using streptavidin beads. Precipitates ( A ) and whole cell lysates (WCL) ( A ′) were analyzed by SDS-PAGE under reducing (R) and non-reducing (NR) conditions followed by immunoblotting (IB). The immunoblots are representative of 3 independent experiments (n = 3). ( B, B′ ) Complementary affinity purification experiment: A tagged version of MEKK4 (MEKK4-SBP) was expressed in HEK293T cells. Cells were treated with H 2 O 2 (100 μM) and lysed at indicated time points. MEKK4-SBP was affinity-purified using streptavidin beads. Precipitates ( B ) and WCL ( B ′) were analyzed by SDS-PAGE under R and NR conditions followed by IB. e.v.: empty vector, PD: pull down, *: endogenous protein, X: unknown protein. The immunoblots are representative of 3 independent experiments (n = 3).

Article Snippet: Jafrac1 was detected with polyclonal goat anti-Prx2 antibody (R&D Systems #AF3489), Drosophila α -tubulin with a monoclonal mouse antibody (Hybridoma Bank AA4.3), Drosophila p-p38 (Thr180/Tyr182) with a monoclonal rabbit antibody (Cell Signaling Technology #4631), Drosophila p38 with a goat polyclonal antibody (Santa Cruz Biotechnology sc-15714) diluted 1:300, myc-tag with a rabbit monoclonal antibody (Cell Signaling Technology #2278), SBP-tag with a mouse monoclonal antibody (Santa Cruz Biotechnology sc-101595), human Prx2 with a rabbit monoclonal antibody (abcam #109367), MEKK4 with a mouse monoclonal antibody (sc-166196), human p-p38 with a mouse monoclonal antibody (BD #6122889) diluted 1:2000, human p38 with a rabbit polyclonal antibody (Cell Signaling Technology #9212) and human β-tubulin with a rabbit monoclonal antibody (Cell Signaling Technology #2128).

Techniques: Affinity Purification, SDS Page, Western Blot, Plasmid Preparation