PC12 cells with indicated treatment and transfections were seeded in a 96-well plate for 48 hours. MTT solution (Dojindo, Tokyo, Japan) was added into each well and incubated for 4 hours. Absorbance at 570 nm was measured by Thermo Multiskan MK3 (Thermo Fisher Scientific Inc, Waltham, MA, USA) followed by incubation with dimethyl sulfoxide. For cell apoptosis, PC12 was harvested, and suspended in binding buffer of ApoDETECT Annexin V-FITC Kit (Thermo Fisher Scientific Inc). The apoptotic cells were analyzed by FACS flow cytometer (Attune, Life Technologies, Darmstadt, Germany) followed by staining with Annexin V-FITC and PI (Thermo Fisher Scientific Inc) according to previous study [15 (link)].
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MTT Assay and Annexin V-FITC/PI Apoptosis Analysis
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Variable analysis
independent variables
- Indicated treatment
- Transfections
- Cell viability (measured by MTT assay)
- Cell apoptosis (measured by Annexin V-FITC and PI staining and flow cytometry)
- PC12 cells
- Incubation time (48 hours for cell viability, unspecified for cell apoptosis)
- Measurement techniques (absorbance at 570 nm for MTT assay, flow cytometry for apoptosis)
- Positive control: Not specified
- Negative control: Not specified
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