Evaluating p16INK4a in Cell Cycle

To observe the effect of p16^INK4a on the cell cycle, CMs were treated with Ad5-cTNT-INK4a or Ad5-cTNT-INK4a RNAi for 48 hours, and control virus groups were established. CMs were digested with 0.25% trypsin without EDTA and centrifuged (1200 rpm, 5 min, 4°C). Then, the cells were washed twice using precooled PBS and resuspended with 70% ethanol. The ethanol suspension was stored overnight in a 4°C refrigerator for cell fixation. Finally, PINase staining buffer (BD Biosciences, USA) was used for dyeing and flow cytometry detection.

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Sun J., Zhou L., Yang T., Deng B., Bao Y., Gu L., Wang H, & Wang L. (2023). P16INK4a Regulates ROS-Related Autophagy and CDK4/6-Mediated Proliferation: A New Target of Myocardial Regeneration Therapy. Oxidative Medicine and Cellular Longevity, 2023, 1696190.

Publication 2023

Buffer Cell Cell cycle Edta Ethanol Flow cytometry Ink4a P16ink4a Rnai Trypsin Virus

Corresponding Organization : Jiangsu Province Hospital

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Variable analysis

independent variables

Treatment with Ad5-cTNT-INK4a
Treatment with Ad5-cTNT-INK4a RNAi

dependent variables

Cell cycle

control variables

Treatment with control virus
Cell digestion with 0.25% trypsin without EDTA
Centrifugation at 1200 rpm for 5 minutes at 4°C
Cell washing with pre-cooled PBS
Cell fixation with 70% ethanol overnight at 4°C
PI/RNase staining buffer for flow cytometry detection

controls

Positive control: Ad5-cTNT-INK4a treatment
Negative control: Ad5-cTNT-INK4a RNAi treatment
Control virus group

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