Research Paper Volume 10, Issue 10 pp 2855—2873

The mitomiR/Bcl-2 axis affects mitochondrial function and autophagic vacuole formation in senescent endothelial cells

Figure 3. Effect of FBS deprivation on young and senescent HUVECs. yHUVECs and sHUVECs were cultured for 48 (48h) or 72 (72h) hours with or without (w/o) FBS. Annexin V positivity and casp-3 activation were analyzed by flow cytometry. (A) Percentage of annexin V-positive cells (left) and ratio of annexin V-positive apoptotic cells among yHUVECs and sHUVECs w/o FBS to their control (with FBS) (right). (B) Percentage of cells with active casp-3 (left); ratio of yHUVECs or sHUVECs w/o FBS with activated casp-3 to control cells (right). (C) Bcl-xL mRNA fold change in yHUVECs and sHUVECs. (D) Western blot and densitometric analysis of Survivin expression in yHUVECs and sHUVECs. (E) Western blot and densitometric analysis of Bcl-2 in yHUVECs and sHUVECs. (F) Percentage of HUVECs showing mPTP opening. Protein expression values are reported as Bcl-2 and Survivin fold change in sHUVECs vs yHUVECs. Data are normalized to β-actin protein. Data are mean ± SD of three independent experiments. * t -test p < 0.05, ** t-test p < 0.01, *** t-test p < 0.001.