Annexin 5 pe 7 aad kit

A549 and NCI-H1299 cells were plated in 60 mm dishes and allowed to attach overnight. Cell apoptosis detection was analyzed using Annexin V-PE/7-AAD kit (catalog KGA1024, KeyGEN). The FACSCalibur flow cytometer (Becton Dickinson) was used to conduct the apoptosis analysis. Data for apoptosis were analyzed using FlowJo7.6 software v7.6.5.

Apoptosis was assessed with the Annexin V-FITC/PI kit (DOJINDO, Beijing, China) with no light for transiently transfected cells, and with the Annexin V-PE/7-AAD kit (KeyGEN BioTECH, Nanjing, China) with green fluorescence for stably transfected cells. Cells were harvested and stained according to the manufacturer's instructions, and then were analyzed by flow cytometry. Data analysis was performed with CELLQuest 3.0 software. The upper right quadrant represents late apoptosis, while the lower right quadrant represents early apoptosis. The calculation of the total apoptotic rate included both the upper and lower right quadrants.

Silica nanoparticles (SiNPs, 637246) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and Turbofect transfection reagent were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Annexin V-PE/7-AAD kit, total protein extraction kit, and BCA protein assay kit were purchased from Nanjing Keygen Biotech Co., Ltd. (Nanjing, Jiangsu, China). DAPI (4′, 6-diamidino-2-phenylindole) was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Anti-GRP78 antibody (C50B12), anti-cleaved Caspase-3 antibody (9664), and anti-mouse and anti-rabbit HRP-linked secondary antibodies (7076P2, 7074P2) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-ERO1α antibody (H00030001-M01) was purchased from Abnova (Taipei, Taiwan). Anti-BCL-2 antibody (ab182858), anti-BCL-xL antibody (ab32370), anti-MCL-1 antibody (ab32087), anti-PUMA antibody (ab33906), anti-BAX antibody (ab32503), anti-BAD antibody (ab32445), and anti-mouse and anti-rabbit fluorescent secondary antibodies (ab150077, ab150115) were purchased from Abcam Ltd. (Cambridge, MA, USA). Anti-CHOP antibody (sc-7351) was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Anti-β-actin antibody (KM9001T) was purchased from Tianjin Sungene Biotechnology Inc. (Tianjin, China).