Fig. 3

HSP70-2 depletion results in apoptosis of breast cancer cells. a Representative SEM images show onset of apoptosis characterized by membrane blebbing, holes and apoptotic bodies formation in shRNA3 and shRNA4 transfected MCF7 and MDA-MB-231 cells compared to NC shRNA transfected cells. Magnification: ×2000, WD = 6 mm, EHT = 20.00 kV. b Flow cytometric analysis shows increased number of MDA-MB-231 cells bound to annexinV when transfected with shRNA3 and shRNA4 compared to NC shRNA transfected cells. c Flow cytometric data demonstrates loss of mitochondrial membrane potential in MDA-MB-231 cells transfected with shRNA3 and shRNA4 compared to NC shRNA transfected cells. d Flow cytometric analysis depicts increased number of MDA-MB-231 cells expressing M30 on their surface when transfected with shRNA3 and shRNA4 compared to NC shRNA transfected cells. e FACS data shows increased DAPI intensity in MDA-MB-231 cells transfected with shRNA3 and shRNA4 compared to NC shRNA. f FACS analysis demonstrates increased apo-BrdU staining in shRNA3 and shRNA4 transfected MDA-MB-231 cells compared to NC shRNA transfected cells. g Western blot analysis shows up-regulation of pro-apoptotic (BAX, BAK, BAD, NOXA, BID, cytochrome-C, PUMA, PARP1, AIF, APAF1, Caspase 3, Caspase 6, Caspase 7, Caspase 9) molecules and down-regulation of anti-apoptotic molecules (BCL2, BCL-x_L, cIAP2, MCL1, Survivin, XIAP, GRP78) in HSP70-2 depleted MDA-MB-231 cells. The three independent experiments were carried out in triplicates