Figure 4.

Continuous long‐term (28 days) everolimus treatment induces drug tolerance in vivo. GFP‐LC3 mice received osmotic minipumps releasing either everolimus (EVR, 1.5 mg·kg⁻¹ per day) or vehicle for a period of 28 days. Prior to killing, mice received an i.p.injection of 100 mg·kg⁻¹ chloroquine. Western blotting was performed to measure the phosphorylation status of Akt1 (Ser⁴⁷³), mTOR (Ser²⁴⁴⁸) and S6rp (Ser) as well as the expression of Cdk2. ACTB was used as a loading control. Representative blots are shown (n = 6 for both groups, *P < 0.05, significantly different from vehicle).