Figure 3
AGR2 knockdown induces cell death. ZR-75-1 cells were collected 96 hours after AGR2 knockdown and analyzed for cell death by measuring ssDNA breaks by using the following: (a) F7-26 staining by FACS analysis, and (b) alterations in mitochondrial membrane potential by determining the ratio of JC-1red to JC-1green and represented as a ratio of the untransfected control (±SD), n = 3. MG132 and CCCP served as apoptosis and depolarization controls, respectively. Cell death was investigated 96 hours after AGR2 knockdown in T47 D cells by using the following: (c) annexin V (AV) and propidium iodide (PI) and gated on normal (AV-/PI-), necrotic (AV-/PI+), early apoptotic (AV+/PI-), and late apoptotic/necrotic (AV+/PI+) cells; and (d) JC-1 staining, as previously described.