UVB-Induced Senescence Evaluation

HaCaT cells were exposed to UVB (100 mJ/cm²) radiation, then treated with CHS for 24 h. For SA-β-Gal (senescence-associated β-galactosidase) staining, the cells were fixed and incubated with a staining solution containing X-gal at 37 °C for two days. For the AO/EB (acridine orange/ethidium bromide) double-staining assay, the cells were stained with a mixture of AO and EB (5 μg/mL) for 10 min in PBS buffer. The MDC (monodansylcadaverine) staining was measured at an excitation wavelength of 335 nm and an emission wavelength of 512 nm. And the fluorescence images were captured using a fluorescence microscope (Zeiss, Oberkochen, Germany).