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Fig. 4 GSH depletion hyperactivates mTORC1 and its repletion protects against aberrant mTORC1 activation. Primary neuronal-glial cerebrocortical cultures were pre-treated with 30 μM dimercaprol for 4 h followed by 100 μM BSO treatment for 20 h in MEM media. 20 nM rapamycin was used as procedural/positive control. mTORC1 activity represented by p-S6/t-S6 was assessed by immunoblotting. No treatment control (NT) represents primary neuronal-glial cerebrocortical cultures treated with plain serum-free MEM media. This control was run to assess basal mTORC1 activity in the cultures and it was not significantly different (P = 0.4767) compared to the vehicle control group. (A) Representative mTORC1 immunoblot, and (B) Blot quantification. Data are represented as mean ± SEM (error bars). *p < 0.05, ****p < 0.0001, ns: not significant versus vehicle control; ββp<0.01 versus 100 μM BSO by one-way ANOVA with Tukey's post-hoc test. n = 2–3/grp. N = 4 experimental replicates.