we studied whether AKT plays a part in TRPC1 mediated neuroprotection connection between AKT and neuroprotection. A decrease buy Linifanib in AKT phosphorylation was observed in PD patient samples, as shown in Figure 5A. Curiously, MPP treatment also significantly decreased AKT1 phosphorylation without affecting overall AKT1 levels in SHSY5Y cells. In addition, over-expression of full length TRPC1, however not TRPC1pm, prevented the decline in AKT phosphorylation seen after MPP treatment. Moreover, quantification of the phospho AKT indicated an approximately 5000-10,000 inhibition of the AKT activity after MPP treatment, that was restored to approximately 75% in cells overexpressing TRPC1 and treated with MPP.. We next examined whether SOCE that is dependent on TRPC1 activates AKT phosphorylation in SH SY5Y cells. Curiously, pro-peptide SH SY5Y cells treated with Tg in the absence of external Ca2 failed to exhibit AKT phosphorylation, suggesting that Ca2 influx through SOCs was required for AKT1 phosphorylation, as Ca2 release from internal ER stores by itself wasn’t adequate to trigger AKT1 phosphorylation. Furthermore, stimulation of TRPC1 by Tg or carbachol considerably improved AKT1 phosphorylation in comparison with control untreated cells. Moreover, addition of SKF 96365 prevented the activation of AKT1 caused by CCh and Tg. We activated SH SY5Y cells with oleyl acetyl glycerol, that is known to activate other TRPC channels and is independent of store depletion, to judge whether other sources of Ca2 trend can also stimulate AKT phosphorylation. Curiously, met inhibitor AKT phosphorylation wasn’t changed upon OAG stimulation, indicating that the effect observed in AKT phosphorylation depends on Ca2 entry via the SOC channel. Additionally, expression of brain-derived neurotrophic factor was also evaluated, since Ca2 entry is famous to stimulate the expression of these factors, that has demonstrated an ability to improve safety of DA cells. Nevertheless, no upsurge in BDNF expression was observed in cells overexpressing TRPC1, indicating that TRPC1 mediated protection is independent of BDNF, as mentioned in Supplemental Figure 6F, bdnf expression was significantly decreased by addition of MPP. We conducted MTT assays, to help measure the role of AKT and TRPC1 in cell survival. MPP treated cells showed a substantial decrease in neuronal survival, which was inhibited by overexpression. Moreover, silencing of AKT1 completely blocked TRPC1 mediated neuroprotection against MPP, showing that AKT1 plays a crucial part in TRPC1 mediated neuroprotection. These strongly claim that TRPC1 mediated Ca2 influx is essential for AKT1 service in SH SY5Y cells, which will be essential for their survival. TRPC1 over-expression shields DA neurons in a in vivo MPTP type of PD. Nothing is known about the function of TRPC1 in an in vivo PD model, while the above strongly suggest the importance of TRPC1 in cellular types of PD.