Fig. 4: Mechanism study of PES for inducing cell death.

a Live/dead staining fluorescence images of B16F10 cells after the different treatments (Control, Laser, MXene, PTT, ES, and PES). The scale bar is 100 μm. b Cell viability of B16F10 cells after the different treatments. P values were calculated by two-tailed t-tests. (n = 3). c, RT-PCR analyses of Caspase-3, GSDME, IL-1β, Bax, c-Jun, Cyt-c expression within B16F10 cells before and after the PES treatments t (n = 4). d The lactate dehydrogenase (LDH) release levels from B16F10 cells after the treatments under different conditions. P values were calculated by two-tailed t-tests. (n = 3). e ATP content change within B16F10 cells under the same cell number after different treatments tested using the ATP assay kit. P values were calculated by two-tailed t-tests. (n = 3). f Bio-TEM images of B16F10 cells before and after the PES treatments. “M” represents mitochondria, “N” represents cell nucleus. g Fluorescence images of ROS within B16F10 cells detected using the DCFH-DA probe after the different treatments. The scale bar is 50 μm. h The fluorescence images of the JC-1 monomer and aggregate, and their merged images within B16F10 cells stained by JC-1 after the different treatments. The scale bar is 50 μm. i Fluorescence images of DNA double-strand breaks within B16F10 cells after the different treatments (corresponding to red fluorescence of γ-H2AX) followed by DAPI staining (corresponding to blue fluorescence). The scale bar is 50 μm. The experiments were repeated for three times with similar results obtained. Data are presented as mean ± SD.