Senescence-associated-β-galactosidase (SA-β-gal) staining for cellular senescence

Jiraporn Kantapan; Siwaphon Paksee; Aphidet Duangya; Padchanee Sangthong; Sittiruk Roytrakul; Sucheewin Krobthong; Wipob Suttana; Nathupakorn Dechsupa

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Senescence-associated-β-galactosidase (SA-β-gal) staining for cellular senescence

JK Jiraporn Kantapan

SP Siwaphon Paksee

AD Aphidet Duangya

PS Padchanee Sangthong

SR Sittiruk Roytrakul

SK Sucheewin Krobthong

WS Wipob Suttana

ND Nathupakorn Dechsupa

This method is extracted from research article: BMC Complement Med Ther, Jul 2021

A radiosensitizer, gallotannin-rich extract from Bouea macrophylla seeds, inhibits radiation-induced epithelial-mesenchymal transition in breast cancer cells

DOI: 10.1186/s12906-021-03363-6

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Cells were seeded into 6-well plates for 24 h to allow attachment before being treated with either MPSE, IR alone, or MPSE combined with 6 Gy IR X-ray. Cells were stained using a senescence cells histochemical staining kit (Millipore Corporation, Bedford, MA, USA) following the manufacturer’s instructions at day 6 after irradiation. Briefly, cells were washed twice by PBS 1× before adding 1× fixation buffer, then incubating at room temperature for 6 min. The fixation buffer was removed, then rinsed off three times with PBS 1×. Cells were stained using a staining mixture and then incubated at 37 °C without CO₂ for 2 h. The cells were observed under a microscope (Nikon, ECLIPSE Ts2, Tokyo, Japan) at a 200× magnification and the percentages of blue-stained cells (senescent cells) were calculated.

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