Annexin 5 fitc pi apoptosis assay kit

Lentivirus-infected cells were sorted based on expression of GFP in successfully infected cells. Apoptotic cells were counted after staining using an Annexin V-FITC/PI Apoptosis Assay Kit (DOJINDO) according to the manufacturer’s instructions. Cells were passed through a 40 µM griddle to discard clustered cells before being analyzed on a BD FACSAria III flow cytometer. Data were processed using the software FlowJo.

The cell counting kit-8 (CCK8) and Annexin V-FITC/PI apoptosis assay kit were purchased from DOJINDO LABORATORIES (Tokyo, Japan). Bepridil (Cat# 4117), KB-R7943 (Cat# 1244), YM-244769 (Cat# 4544), SN-6 (Cat# 2184), and [1,2-bis(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA-AM)] (Cat# 2787) were from Tocris Bioscience (Minneapolis, USA). The BRAF inhibitor vemurafenib (Cat# HY-12057) were from MedChemExpress (Monmouth Junction, NJ, USA). These compounds were dissolved in dimethyl sulfoxide (DMSO) to make stock solution. Propidium iodide (PI) and Ribonuclease A were from Sigma Aldrich (St. Louis, MO, USA). Cell-permeable Ca²⁺ indicator Fluo-4 AM was from Invitrogen Life Technologies (Waltham, MA, USA). The primary antibody for NCX1 isoform (Cat# ab177952) was from Abcam (Cambridge, MA, USA); anti-NCX2 (Cat# ANX-012) and NCX3 (Cat# ANX-013) antibodies were from Alomone Labs (Jerusalem, Israel). Mouse α-tubulin antibody (Cat# 2144) was from Cell Signaling Technology (Boston, MA, USA).