Fig 1: A summary figure showing mechanism of ouabain cytotoxicity towards keratinocytes. Low doses of ouabain cause no detectable inhibition of the Na + /K + pump activity, but inhibits activation of Akt and mTOR, which leads to upregulation of beclin. Beclin forms a complex with SLC7A11 and directly reducing its protein expression, decreasing cystine uptake. Consequently, intracellular GSH synthesis declines, causing excessive oxidative stress and ultimate cell death
Fig 2: Ouabain downregulated SLC7A11 protein via Akt/mTOR/beclin axis. The total and phosphorylated levels of a Akt and b mTOR were determined by a colorimetric method using the Phospho-Akt/mTOR ELISA kit. Beclin (c) and SLC7A11 (d) protein levels were determined by Western blots. Cells were treated with 200 nM ouabain for 12–24 h, with or without MHY1485 pretreatment (10 μM, 30 min). e Cystine uptake was measured using a commercialized kit. *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant
Supplier Page from Abcam for Phospho-AKT / GSK3 beta / mTor ELISA kit