Fig. 5.

Mitochondria membrane potential (Δψm) loss and activation of apoptotic signaling. (A, B) Cells were treated with vehicle (control), gluRDVs, free Dox, Dox-gluRDVs at equally higher concentration of Dox (10 μM) or FCCP (100 μM as positive treatment) after 8 h of incubation and then stained with Rh-123 for microscopic observation (A) and flow cytometry (B). (A) Rh-123 staining for mitochondrial membrane potential. Reduced Rh-123 fluorescence (green) was shown in Dox-gluRDVs-treated cells. Hoechst33342 staining for nucleus (blue). Scale bar: 10 μm. (B) Flow cytometry quantification of Δψm presented by mean fluorescence intensity (MFI) of Rh-123 (p-values: * < 0.05 as compared with control; # < 0.05 as compared with free Dox). (C) Cells were treated with vehicle (control), gluRDVs, free Dox or Dox-gluRDVs at equally lower concentration of Dox (0.3 μM) after 24 h of incubation and then processed for western blot analysis of ERK phosphorylation and active caspase-3. GAPDH as internal control.