Evaluation of Cell Injury Signaling

Soluble dsDNAs in the supernatant were evaluated by dsDNA Assay kit (see Supplementary Materials) [35 (link)]. Cell injury and the associated signal were evaluated by TUNEL assay [35 (link)] and immunostaining of NF-κB p65 subunit and STAT3 using anti-NFκB antibody (Abcam, Cambridge, UK) and anti-STAT3 antibody (Thermo Fisher Scientific), respectively, after fixation with 4% paraformaldehyde. The positive areas of TUNEL staining and immunostaining were measured using the ImageJ software as described [38 (link)]. The data were expressed as the mean ± standard error of the mean (SEM) from three independent experiments.