Anti cd338

The immunophenotype of MT-CHC01 cells was determined by flow cytometric analysis. Cells were washed in 1× PBS containing 0.1 % bovine serum albumin (BSA, Sigma-Aldrich, Saint Louis, MO, USA) and 0.01 % sodium-azide. For cell permeabilization, when requested, the Fix and Perm reagent (BD Italia) was used following the manufacturer’s instructions. The following antibodies were used: fluorescein isothiocyanate (FITC)-conjugated mouse anti-CK7 and anti-CK19 (Abcam Cambridge, UK) and CD44 (BD Bioscience Europe), allophycocyanin (APC)-conjugated mouse mAbs anti-EPCAM (BD), anti-CD34 and anti-CD133 (Miltenyi BiotecS.r.l., Italy), phycoerytrin-conjugated (PE) anti-CD24, anti-CXCR4, anti-Oct3/4, anti-FOXA1/2, anti-PDX1 (all from BD), and anti-CD338 (R&D Systems, Inc. Minneapolis, MN), PerCP-Cy 5.5-conjugated anti-SOX2/17, Alexafluor 647-conjugated anti-Nanog and anti-Stro1 (BD), and Alexafluor 488-conjugated anti-PAX6 (BD).

Immunophenotyping of 82.3 cells was performed by flow cytometric analysis. Cells were washed in 1× PBS containing 0.1% bovine serum albumin (BSA, Sigma-Aldrich, Saint Louis, MO, USA) and 0.01% sodium azide. For intracellular markers, cells were permeabilized with fix and perm reagent (BD Italia) following the manufacturer’s instructions. The following antibodies were used: FITC (Fluorescein isothiocyanate) conjugated mouse mAbs anti-CK7 and anti-CK19 (Abcam, Cambridge, UK) and anti-CD44 (BD Bioscience, San Jose, CA, USA), APC-(allophycocyanin) conjugated mouse mAbs anti-EPCAM and anti-AFP (BD Bioscience Europe), anti-CD34 and anti-CD133 (Miltenyi Biotec S.r.l., Bologna, Italy), PE (phycoerytrin) conjugated mAbs anti-CD24, anti-CXCR4, anti-Oct3/4, anti-FOXA1/2, anti-PDX1, (all from BD), anti-CD338 (R&D Systems, Inc., Minneapolis, MN, USA), PerCP-Cy 5.5 conjugated mAbs anti-SOX2/17, Alexa Fluor 647 conjugated mAbs anti-Nanog, anti-Stro1, and Alexa Fluor 488 conjugated mAbs anti-PAX6 (BD Bioscience Europe).