Senescence-Associated β-Galactosidase Assay

SA-β-Gal staining was performed as previously described (29 (link)). Briefly, cultured cells were washed twice with PBS and fixed at room temperature for 5 min in 2% formaldehyde and 0.2% glutaraldehyde. Fixed cells were stained with SA-β-Gal staining solution (40 mM citric acid/Na phosphate buffer, 5 mM K₄[Fe(CN)₆], 5 mM K₃[Fe(CN)₆], 150 mM NaCl, 2 mM MgCl₂, 1 mg/ml X-gal) at 37°C overnight, and images were captured using a microscope digital camera (Olympus). The percentage of SA-β-Gal-positive cells were calculated with ImageJ.

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Wu Z., Shi Y., Lu M., Song M., Yu Z., Wang J., Wang S., Ren J., Yang Y.G., Liu G.H., Zhang W., Ci W, & Qu J. (2020). METTL3 counteracts premature aging via m6A-dependent stabilization of MIS12 mRNA. Nucleic Acids Research, 48(19), 11083-11096.

Publication 2020

microscope digital camera

Buffer Camera Citric acid Cultured cells Formaldehyde Glutaraldehyde Mgcl2 Microscope Nacl Phosphate X gal

Corresponding Organization : Capital Medical University

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Variable analysis

independent variables

SA-β-Gal staining

dependent variables

Percentage of SA-β-Gal-positive cells

control variables

Cell culture conditions
Fixation time (5 min)
Fixation temperature (room temperature)
Fixation solution (2% formaldehyde and 0.2% glutaraldehyde)
Staining solution (40 mM citric acid/Na phosphate buffer, 5 mM K4[Fe(CN)6], 5 mM K3[Fe(CN)6], 150 mM NaCl, 2 mM MgCl2, 1 mg/ml X-gal)
Staining temperature (37°C)
Staining duration (overnight)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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