Figure 5.

A) Flow cytometric analysis of time-dependent change in intracellular Ca²⁺ represented by Fluo-3AM fluorescence intensity. Raji cells pretreated with Fab′-MORF1 and loaded with Fluo-3AM were excited at 488 nm and emission was measured at 530 nm on flow cytometry. A baseline was obtained for 60 s before stimulation with P-(MORF2)_x crosslinking (indicated by black arrow). B) Confocal microscopy visualization of Raji cells stained with Fluo-3AM (green fluorescence) after DFMT treatment for 1 h. C) Apoptosis evaluation of DFMT with or without various inhibitors by Annexin V/PI method. To determine the extracellular calcium influx or intracellular calcium release, extracellular calcium free condition was created by adding 10 × 10⁻³ M EGTA to 1640 medium. To deplete cholesterol and inhibit CD20 crosslinking, cells were preincubated 1 h at 37 °C in the presence of β-CD.