Circadian Regulator circ-ATXN2 Modulates Apoptosis

To observe the effects of circ-ATXN2 overexpression on cell death, CoCl₂ was used to create apoptotic models of the cells. Vector-GFP- and circ-ATXN2–GFP-expressing cells were seeded at 1 × 10⁵ cells per well in 6-well plates and incubated with 500 μM CoCl₂ for 24 h. Annexin V-APC/PI apoptosis kit (MULTI SCIENCES) was used to determine cell apoptosis. Cells were collected in 500 μl of binding buffer with 3 μl Annexin V-APC and 3 μl PI for 20 min at room temperature in the dark. Apoptotic cells were detected using a flow cytometer (DxFLEX, Beckman Coulter, United States) (He et al., 2021 (link); Song et al., 2021 (link)).