Figure 3
From: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells
69-kDa Ku86v in MM cell lines is not the result of post-translational modification by N-linked deglycosylation. Freshly obtained cell lysates (30 μg/sample) from the SGH-MM5 MM cell line were incubated with or without Endo H (500 U/reaction) for 2 hrs or 24 hrs (A); or with varying amounts (0, 1, 2, 3, 4 or 5 μL) of Endo H (500 U/μL) for 20 hrs (B; positive control); and then resolved in a 12.5% SDS-PAGE gel, and immunoblotted using S10B1 anti-Ku86 (A), or anti-heavy chain of the human MHC class I protein mAbs (B). Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Experiments were performed in triplicate.