Expression of NKG2D-L, ERp5, and ADAM10 in LN MSCs. LN MSCs were obtained from cHL patients (n = 8, 6 NS, 2 MC). (A) Staining with specific mAbs recognizing HLA-I, SH3/CD73a, CD90, SH2/CD105 (Ai), PH4, collagen, vimentin, and TG (Aiii), followed by PE-conjugated anti–isotype specific GAM (black lines). Gray lines indicate negative control staining with isotype-matched irrelevant mAbs followed by PE-GAM. Samples were run on a CyAnADP flow cytometer, and results are expressed as log red MFI (a.u. indicates arbitrary units) versus number of cells; in each histogram are indicated the MFI and the percentage of positive cells. (Aii,iv) MFI ratio between MFI of positive cells for each marker and MFI of the negative control. Data are mean ± SD from 8 LN MSCs. (B) Surface (Bi) or cytoplasmic (Biii) staining of LN MSCs with anti–MIC-A or anti-ULBP1 to ULBP4 mAbs. For cytoplasmic staining, cells were fixed in 3% paraformaldehyde and permeabilized with1% Nonidet-P40 in PBS, before mAb incubation. Results are expressed as mean red fluorescence intensity; in each histogram are shown the MFI and the percentage of positive cells. (Bii,iv) MFI ratio as in panel A. Data are mean ± SD from 8 LN MSCs. (C) Surface (Ci) and cytoplasmic (Ciii) staining of LN MSCs with the anti–ERp5 rabbit polyclonal antibody or anti–ADAM10 mAb, followed by FITC-goat anti–rabbit (GAR) or anti–isotype GAM antisera (black lines). Gray lines indicate negative control staining with unrelated irrelevant Abs. Results are expressed as log green mean fluorescence intensity; in each histogram are indicated the MFI and the percentage of positive cells. (Cii,iv) MFI ratio: ratio between MFI of a given marker and the negative control. Data are mean ± SD from 8 LN MSCs. (D) Soluble MIC-A, ULBP2, and ULBP3 were measured in the supernatant of cultured LN MSCs by ELISA, referred to a standard curve, and expressed as nanograms per milliliter. Data are mean ± SD from 8 LN MSCs.