Fig 2: mTOR signalling. (A) mTOR phosphorylation relative to total mTOR expression. (B) Raptor phosphorylation relative to total Raptor expression. (C) Rictor phosphorylation relative to total Rictor expression, following 16 h with no treatment or with TGFß1 (1 ng/ml or 5 ng/ml) and 30 min insulin stimulation (0 nM or 100 nM). Data are reported as Mean ± SEM. AU, arbitrary units (defined as band density values). Phospho (P) and Total (T). Healthy: N = 5 participants (clear bars), PCOS: N= 5 participants (filled bars).

Fig 3: Mean and standard deviation of the relative expression of the genes coding for the same of the proteins that constitute the mTOR complexes, normalised to Saline/Withdrawal 1 (VhSA) values. The rats that self-administered heroin showed higher mRNA levels of Rictor than their saline-treated controls after 1 day of withdrawal (F1,28 = 5.094, p = 0.032, ?2 = 0.182). Also, among the groups that self-administered saline solution, the rats that underwent 30 days of withdrawal showed higher Rictor mRNA levels than the rats that only had one day of withdrawal (F1,28 = 7.104, p = 0.013, ?2 = 0.254). *Significant effect of the simple effect analyses (p < 0.05)

Fig 4: Representative western blot images and densitometry bar graphs normalised to total protein loaded (Rictor and Phospho-Rictor (thr1135) or to Rictor levels (p-Rictor/Rictor)). The rats that self-administered heroin showed higher phosphorylation rates at thr1135 than their saline-treated controls (F1,22 = 5.247, p = 0.032, ?2 = 0.155). $Significant effect of the treatment factor (p < 0.05)

Fig 5: AGR2 upregulation during emergence induces AKT and RICTOR activation via phosphorylation. During CIS escape, AGR2 expression is induced and allows the phosphorylation of both AKT and RICTOR. The two pathways are known to contribute to cell proliferation. RICTOR activation might also be induced by the extracellular AGR2. AGR2, anterior gradient 2; CIS, chemotherapy-induced senescence; mTORC, mammalian target of rapamycin complex; eAGR2, extracellular AGR2.