Abrogation of the treasure induced upregulation of IL 1Ra mRNA in fMCNs by wortmannin and LY294002 shows the participation of PI3 K in neuronal upregulation of IL Canagliflozin 1Ra. This is further verified by IL 1Ra immunofluorescence in fMCNs. Involvement of Akt in gem mediated upregulation of IL 1Ra in fMCNs Since PI3 K is known to activate the downstream kinase Akt, we investigated if Akt was involved with gem induced upregulation of IL 1Ra. First, we examined if gem alone was effective at causing the activation of Akt by monitoring levels of phosphorylated Akt using antibodies against Akt p Ser473. The amount of total Akt was unchanged, while diamond time dependently induced the phosphorylation of Akt. Densitometric analysis of r Akt indicated that gem was seen at 15, 30 and 60 min of gem treatment and that considerable elevation of pAkt was capable of causing the phosphorylation of Akt since 5 min. We immunostained fMCNs for p Akt and MAP 2, to further ensure the activation of Akt. Again, we noticed an increase in g Akt at 15 and 30 min of treasure coverage relative to control. These results suggest diamond alone is effective at causing the activation of Akt in fMCNs. Next, to observe the involvement of Akt in gem induced upregulation pro-protein of IL 1Ra, we used Akt i, a specific inhibitor of Akt. RT PCR and real-time PCR analyses indicate an increase in IL 1Ra mRNA expression in the presence of jewel alone. This upsurge in IL 1Ra mRNA was abrogated when fMCNs were preincubated with Akt i. To further confirm this statement, we conducted double brand immunofluorescence for MAP 2 and IL 1Ra. Akt i substantially restricted diamond induced up-regulation of IL 1Ra in fMCNs, as evident from figure 4F. These results suggest an obligatory function for Akt in the diamond mediated up-regulation of IL 1Ra in neurons. CREB is needed for gem to induce IL 1Ra expression Next we examined ALK inhibitor mechanisms where PI3 K Akt pathway coupled IL Ra up-regulation in gem treated neurons. Upon analysis of the IL 1Ra promoter using MatInspector, binding sites were observed by us for many transcription components including one consensus cAMP response element nearby the transcriptional start site. Moreover, CREB plays numerous roles in health and success. Therefore, we were prompted to investigate if diamond needed CREB for the transcription of IL 1Ra in nerves. First, we examined if treasure alone induced the activation of CREB in neurons by checking levels of phosphorylated CREB, DNA binding activity by EMSA and transcriptional activity utilizing a luciferase reporter construct. Gem alone induced the phosphorylation of CREB as indicated by Western blot and immunofluorescence analyses. On the other hand, we did not see any significant change in the degree of total CREB. Next we examined the DNA binding activity of CREB. Diamond treatment induced a slower migrating band, that has been supershifted by antibody against CREB, but not control IgG, confirming the presence of CREB in the protein nucleic acid complex, as observed in figure 5D.