the accumulation of intracellular ROS is related to cell death induced by poisonous heavy metals, this study investigated whether NaF induced intracellular ROS accumulation in mESCs. NaF mediated reduction of stability occurred at 2 mM NaF after Cyclopamine 11-deoxojervine 24 h incubation set alongside the untreated control cells. Almost complete inhibition of viability was observed once the cells were exposed to over 4 mM NaF for 24 h or 2 mM NaF for 72 h. NaF inhibited DNA synthesis in a dose dependent manner. Managing the cells with 3 and 5 mM NaF for 24 h decreased TdR uptake levels by 81 half an hour and 44 six months, respectively, set alongside the non treated get a grip on. Cell cycle analysis unveiled that NaF treatment generated cell population migration in to the sub G1 and G2/M phases having a concomitant decrease of cells in the S phase. Subsequently, the levels of cyclin dependent kinase 2, cyclin E, and proliferating cell nuclear antigen were analyzed by western blot analysis. NaF treatment didn’t affect PCNA and CDK2 protein levels however it markedly reduced cyclin E levels. Flow cytometric analysis after PI staining showed that the cell population in the sub G1 cycle of cell cycle progression, which shows apoptotic cell death, improved after treatment Retroperitoneal lymph node dissection with NaF in a dose dependent fashion. FITC annexin V/PI discoloration tests also unveiled that cell populations demonstrating high PI and high FITC and low PI and high FITC indicators risen to 17. Five hundred and 24. Six months, respectively, after exposing the cells to 5 mM NaF for 24 h as compared to the untreated control level of 2. 0.5-1kg. Figure 3B shows a significant increase in the number of apoptotic cells based on NaF awareness, although there is also a moderate increase in necrotic cells as indicated by the large PI and low FITC signals. NaF mediated apoptosis was supported by results from ELISA Bicalutamide solubility based assays, where NaF treatment induced a dose-dependent increase in DNA strand breaks. Additionally, exposure of mESCs to NaF led to an increase of poly polymerase cleavage and a marked decrease of Akt1 protein levels. Flow cytometric analysis unmasked that NaF treatment increased ROS levels inside the cells in a dose dependent manner. This finding was supported by ESR signals showing the dose-dependent increase of hydroxyl radicals in NaF treated mESCs. Eventually, the results of N acetyl cysteine, catalase, superoxide dismutase, and apocynin anti-oxidants on viability in NaF subjected mESCs were determined. Pre-treatment with 2,500 U/ml CAT, however not with other antioxidants, showed a significant inhibition in the NaF mediated reduction of cell viability. MESCs were subjected to different concentrations of NaF in the absence and presence of 500 and 2,500 U/ml CAT for 24 h, to better comprehend the effects of CAT. As shown in Figure 4D, treating cells with 500 U/ml CAT showed mild defense against NaF caused accumulation only once the cells were exposed to 2 mM NaF, although treatment with 2,500 U/ml markedly inhibited the NaF mediated reduction in cell viability in the exposed NaF concentrations.