Fig. 2
From: CYD0281, a Bcl-2 BH4 domain antagonist, inhibits tumor angiogenesis and breast cancer tumor growth
CYD0281 promotes HUVECs cell apoptosis by the exposure of the Bcl-2 BH3 domain. A Cell viability of HUVECs at 48 h. The exposure of the BH3 domain of Bcl-2 detected by immunofluorescence (IF) staining using anti-Bcl-2/BH3 domain antibody (B), and flow cytometry analysis of cell apoptosis (C) in HUVECs treated with BDA-366 and CYD0281 at the IC50 concentration. HUVECs were treated with CYD0281 at the IC50 concentration for 48 h, mitochondrial dysfunction was analyzed by Mito-Tracker Red CMXRos staining (D), Cyt c release (E) and PARP cleavage (F) were analyzed by western blotting assay, and Bcl-2 associated Bax or Bim was analyzed by co-IP using Bcl-2 antibody (G). H The expression of Bcl-2 in HUVECs was inhibited using siRNAs (1, 2, and 3) and analyzed by western blotting assay. I Flow cytometry analysis of cell apoptosis in Bcl-2-silenced HUVECs that were treated with CYD0281 (n = 3). Significant effect compared to the DMSO group: *P < 0.05 and ***P < 0.001. ns: no significant differences. Scale bars: 50 μm