Journal: Journal of Cellular and Molecular Medicine

Article Title: The endogenous danger signals HSP70 and MICA cooperate in the activation of cytotoxic effector functions of NK cells

doi: 10.1111/j.1582-4934.2008.00677.x

Figure Lengend Snippet: L-MICA cell react with human NKG2D and MICA/B-specific antibodies. The expression of MICA and HSP70 was analysed by flow cytometry on MICA-transfected L-MICA and L-con cells. MICA cell surface expression is shown by staining with mAb against human MICA and MICB (BAMO3 and IIIC1) and by binding of a recombinant human NKG2D-Fc fusion protein. HSP70 cell surface expression was analysed using the mAb cmHsp70.1. Staining with the respective primary reagent (solid line) and FITC-labelled secondary reagent only (dashed line) is shown together with unstained cells (dotted line).

Article Snippet: Furthermore, the MICA specific mAb AMO1 (mouse IgG 1 , Bamomab) and MICB specific mAb BMO2 (mouse IgG 2a , Bamomab) were used.

Techniques: Expressing, Flow Cytometry, Transfection, Staining, Binding Assay, Recombinant

Journal: Journal of Cellular and Molecular Medicine

Article Title: The endogenous danger signals HSP70 and MICA cooperate in the activation of cytotoxic effector functions of NK cells

doi: 10.1111/j.1582-4934.2008.00677.x

Figure Lengend Snippet: Induction of NKG2D ligands MICA/B on human melanoma cells. (A) Ge-con and Ge-Hsp70 cells were analysed by flow cytometry for cell surface expression of NKG2D ligands (human NKG2D-IgG-Fc fusion protein), MICA/B (mAb BAMO1), MICA (mAb AMO1), MICB (mAb BMO1) and HSP70 (mAb cmHsp70.1). The cells were either cultured under standard conditions (co) or exposed to 10 μM SAHA for 20 hrs before the test. The mean percentage + S.D. of cells showing cell surface expression and the mean + S.D. of the specific MFI (MFI for specific staining minus MFI for the secondary reagent only) are shown. The number of independent measurements is given in the upper panel above the bars. (B) Ge-con and Ge-Hsp70 cells were analysed by flow cytometry for cell surface expression of ULBP1, ULBP2, and ULBP3 using specific mAbs. The cells were either cultured under standard conditions (co) or exposed to 10 μM SAHA for 20 hrs before the test. The mean percentage + S.D. of cells showing cell surface expression and the mean + S.D. of the specific MFI (MFI for specific staining minus MFI for the secondary reagent only) are shown as determined in three to four experiments.

Article Snippet: Furthermore, the MICA specific mAb AMO1 (mouse IgG 1 , Bamomab) and MICB specific mAb BMO2 (mouse IgG 2a , Bamomab) were used.

Techniques: Flow Cytometry, Expressing, Cell Culture, Staining