Lenalidomide

Lenalidomide was purchased from (BioVision Inc.), Pomalidomide and Phthalimide were purchased from (Sigma-Aldrich). These drugs were dissolved in dimethylsulphoxide (DMSO) and stored at −20°C until use. The final concentration of DMSO in all experiments was < 0.1%.
The following monoclonal antibodies (mAbs) were used for immunostaining or as blocking Abs: anti-MICA (MAB159227), anti-MICB (MAB236511), anti-ULBP-1 (MAB170818), anti-ULBP-2/5/6 (MAB165903), anti-ULBP-3 (MAB166510) and anti-NKG2D (MAB149810) from (R&D System), anti-PVR/CD155 (SKII.4) kindly provided by Prof. M. Colonna (Washington University, St Louis, MO), anti-CD56 (C218) mAb was provided by Dr. A. Moretta (University of Genoa, Genoa, Italy), anti-DNAM-1 (DX11) from (Serotec), APC Goat anti-mouse IgG (Poly4053), anti-CD3/APC (HIT3a), anti-CD56/PE (HCD56), mouse IgG1/FITC, /PE or /APC isotype control (MOPC-21) were purchased from (BioLegend). Anti-CD107a/FITC (H4A3), anti-CD138-FITC (M15) and anti-CD38-APC (HIT2) were purchased from (BD Biosciences).

The bromodomain inhibitor JQ1 and Bortezomib (PS-341) were purchased from Selleckchem.Com (Munich, Germany). Lenalidomide was purchased from BioVision Inc. (Milpitas, California USA). The hetero-bifunctional PROTAC (Proteolysis Targeting Chimera) ARV-825, was purchased from Chemieteck (Indianapolis, IN, USA). These drugs were dissolved in dimethylsulphoxide (DMSO) and stored at −20 °C until use. The final concentration of DMSO in all experiments was < 0.1%. Roscovitine a potent and selective inhibitor of cyclin-dependent kinases and the antibiotic Doxycycline hydrate (used to induce SKO-007(J3)/shMEIS2-Tet cells), Melphalan and 17-AAG (17-allylaminogeldanamycin) were purchased from Sigma-Aldrich S.r.l. (Milan, IT). The following monoclonal antibodies (mAbs) were used for immunostaining or as blocking Abs: anti-MICA (MAB159227) was purchased from R&D System (Minneapolis, MN), anti-PVR/CD155 (SKII.4) kindly provided by Prof. M. Colonna (Washington University, St Louis, MO), APC Goat anti-mouse IgG (Poly4053), was purchased from BioLegend (San Diego, CA). Anti-CD138-FITC (M15) and anti-CD38-APC (HIT2) were purchased from BD Biosciences (San Jose, CA).

The human K562, NK‐92, and HEK293T cell lines were purchased from ATTC and were maintained at 37°C and 5% CO₂ in medium supplemented with 10% FBS. Note that 200 U/ml human recombinant IL‐2 was added for NK‐92 cells. All cell lines were mycoplasma‐free (Mycoplasma Test Kit, Biological Industries).
Healthy donors’ peripheral blood mononuclear cells (PBMCs) were isolated by Lymphoprep (Nycomed) gradient centrifugation. NK cells were negatively selected from PBMCs using MACS human NK cell isolation kit (Miltenyi Biotec).
Lenalidomide was purchased from BioVision, Inc. (Phthalimide, Wiskostatin), and NSC23766 and MLN4924 were purchased from Merck Life Science. Homo‐PROTAC cereblon degrader 1 was purchased from Selleckchem. The following monoclonal antibodies (mAbs) were used for immunostaining: anti‐CD3/FITC, anti‐CD56/PE, anti‐CXCR4/APC, anti‐GZM‐B/FITC and anti‐GZM‐B/APC, anti‐total LFA1/APC (BD Biosciences), anti‐LFA1 clone 24 CD11‐CD18 (Hycult Biotech), and APC Goat anti‐mouse IgG (Jackson Immunoresearch Laboratories). Phalloidin‐FITC was purchased from Thermo Fisher Scientific (Waltham).
Human recombinant cytokines IL‐2 and CXCL12 were purchased from Peprotech EC.