Journal: Arthritis Research & Therapy

Article Title: Tumour necrosis factor alpha promotes secretion of 14-3-3η by inducing necroptosis in macrophages

doi: 10.1186/s13075-020-2110-9

Figure Lengend Snippet: 14-3-3η is expressed in CD68 + cells but not in CD4 + T cells from the synovium of RA patients. Synovium specimens of patients with RA ( n = 3) were analysed by IF using specific antibodies against 14-3-3η and CD68 ( a ) or CD4 ( b ). c Lung tissue specimens of patients with RA ( n = 3) were analysed using specific antibodies against 14-3-3η and CD68. Representative images of three independent experiments are shown. Scale bar, 50 μm

Article Snippet: The following antibodies were purchased and used for immunohistochemistry (IHC), immunofluorescence (IF), immunocytochemistry (ICC), and immunoblotting (IB) experiments: mouse monoclonal antibodies against 14-3-3η (1:200; AAM-11-0003, Augurex, Vancouver, BC, Canada) and 14-3-3γ (1:200; sc-731, Santa Cruz Biotechnology, Dallas, TX) for IHC, CD4 (1:200; ab67001, Abcam, Cambridge, MA) for IF, CD55 (1:200; ab1422, Abcam) for IF, 14-3-3η (1:200; NBP1-92691SS, Novus Biologicals, Centennial, CO) and phospho-Akt (1:200; Ser473) (#9271 Cell Signaling Technology, Danvers, MA) for ICC, and β-actin (1:10,000; A1978, Sigma-Aldrich, St. Louis, MO) for IB; rabbit monoclonal antibodies against CD68 (1:200; 11192-RP02, Sino Biological Inc., Wayne, PA), PAD4 (1:200; 17373-1, ProteinTech, Rosemont, IL), and phospho-RIP3 (Ser227) (1:200; 93654S, CST) for IF and ICC, and MLKL (1:2000; phospho S358, ab187091, Abcam) for IB; rabbit polyclonal antibodies against MLKL (1:2000; ab194699, Abcam) for IB.

Techniques:

Journal: Arthritis Research & Therapy

Article Title: Tumour necrosis factor alpha promotes secretion of 14-3-3η by inducing necroptosis in macrophages

doi: 10.1186/s13075-020-2110-9

Figure Lengend Snippet: PAD4 is localized in CD68 + cells and CD55 + cells in the RA synovium. Synovium ( n = 3) specimens of patients with RA were assessed by IF using primary antibodies against PAD4 and 14-3-3η ( a ), PAD4 and CD68 ( b ), 14-3-3η and CD55 ( c ), or PAD4 and CD55 ( d ) and by FITC or rhodamine-labelled secondary antibodies. Representative images from three independent experiments are shown. Scale bar, 50 μm

Article Snippet: The following antibodies were purchased and used for immunohistochemistry (IHC), immunofluorescence (IF), immunocytochemistry (ICC), and immunoblotting (IB) experiments: mouse monoclonal antibodies against 14-3-3η (1:200; AAM-11-0003, Augurex, Vancouver, BC, Canada) and 14-3-3γ (1:200; sc-731, Santa Cruz Biotechnology, Dallas, TX) for IHC, CD4 (1:200; ab67001, Abcam, Cambridge, MA) for IF, CD55 (1:200; ab1422, Abcam) for IF, 14-3-3η (1:200; NBP1-92691SS, Novus Biologicals, Centennial, CO) and phospho-Akt (1:200; Ser473) (#9271 Cell Signaling Technology, Danvers, MA) for ICC, and β-actin (1:10,000; A1978, Sigma-Aldrich, St. Louis, MO) for IB; rabbit monoclonal antibodies against CD68 (1:200; 11192-RP02, Sino Biological Inc., Wayne, PA), PAD4 (1:200; 17373-1, ProteinTech, Rosemont, IL), and phospho-RIP3 (Ser227) (1:200; 93654S, CST) for IF and ICC, and MLKL (1:2000; phospho S358, ab187091, Abcam) for IB; rabbit polyclonal antibodies against MLKL (1:2000; ab194699, Abcam) for IB.

Techniques:

Journal: The EMBO Journal

Article Title: A G1‐like state allows HIV ‐1 to bypass SAMHD 1 restriction in macrophages

doi: 10.15252/embj.201696025

Figure Lengend Snippet: A, B Stimulated or unstimulated MDM were stained for classical macrophage markers (CD68, CD14) and M1 and M2 macrophage markers (CD163, CD80, CD86 and CD40) and analysed by FACS.

Article Snippet: Antibodies used were as follows: anti‐cdc2 (Cell Signaling Technology, Beverly, MA, USA); anti‐CDK2 (H‐298, Santa Cruz Biotechnology); anti‐pCDK2(Thr160) (Bioss Inc., MA, USA); anti‐CDK4 (DCS156, Cell Signaling Technology); anti‐CDK6 (B‐10, Santa Cruz Biotechnology); anti‐SAMHD1 (ab67820, Abcam, UK), beta‐actin (ab6276, abcam, UK); mouse anti‐MCM2 (BM‐28, BD Biosciences, UK); and rabbit anti‐MCM2 (SP85) from Sigma; pSAMHD1 (a kind gift from M. Benkirane) and ProSci (Poway, CA, USA); anti‐Geminin (NCL‐L‐Geminin, Leica); anti‐mouse F4/80 and CD11b (kind gift from S. Yona, UCL); anti‐human CD68, CD14, CD163, CD80, CD86, CD40 (kind gift from M. Noursadeghi).

Techniques: Staining