The current results also indicate that hydroxyl radicals are the immediate mediator of NaF mediated cell death, as evidenced by the dose-dependent increase in ESR sign and DCF fluorescence and potent c-Met inhibitor the CAT mediated prevention of cell toxicity in NaF treated mESCs. These data are also consistent with previous findings, in which hydroxyl radicals were proved to be the key harmful radicals in mycotoxin or rock exposed cells. Cytoplasmic release of cytochrome c and its complex development with Apaf 1 and procaspase 9 stimulates executive caspase 3. In the present study, NaF induced a marked cleavage of PARP in mESCs. NaF mediated decrease in cell viability was also suppressed by treatment with a pot caspase inhibitor. These results support strongly the effort of the caspase mediated process in NaF mediated apoptosis in mESCs. More over, our results suggest that the lower in Akt levels relates to a NaFmediated reduction of cell viability, though more descriptive tests to explain the position of Akt in NaF uncovered mESCs will undoubtedly be required. Collectively, the mitochondrial and caspasemediated signaling associated with intracellular ROS accumulation generally seems to Cellular differentiation be engaged in NaF mediated apoptosis. A few studies have suggested the involvement of the JNK pathway in fluoride induced apoptosis. Fluoride publicity at 2 to 10 mM caused prolonged phosphorylation of JNK in MDPC 23 odontoblast like cells. Long-term fluorosis increased g JNK levels in rat brains, which will be just like the outcome of SH SY5Y cells treated with extortionate fluoride. These accounts declare that over exposure to exorbitant fluoride could activate the JNK pathway. There is also considerable evidence that GADD45 comes with an important part in the induction of apoptosis, in which its transcription and function are managed either by JNK1 or JNK2. In a previous study, cadmium improved the production of GADD45 Cyclopamine 11-deoxojervine in JB6 Cl41 cells and this was suppressed by its pharmacological inhibitor or si JNK transfection. In parallel with this report, NaF treatment increased the induction of GADD45 in an amount and time dependent manner and this effect was prevented by a JNK specific inhibitor. On the other hand, NaFmediated MMP damage was inhibited by PFT or CAT, however not by SP600125. More, NaFmediated ROS accumulation was restricted only by CAT in the place of by JNK or p53 inhibitors. These results claim that p53 mediated signaling and JNK GADD45 is crucial for NaF mediated apoptosis in mESCs, where ROS act as the most crucial upstream mediator. Intracellular calcium ions can play vital roles in fluoride induced apoptosis. Intracellular calcium homeostasis can be critical for maintaining cellular functions in reaction to additional and/or endogenous stimuli. Similarly, cadmium elevated intracellular calcium levels and then mediated apoptosis. However, the present study unmasked the other result, for the reason that treatment with calcium-channel blockers didn’t restrict NaFmediated reduction in cell viability, relatively BAPTA AM assisted the NaF mediated harmful effects.