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Fig. 6 | Cell Communication and Signaling

Fig. 6

From: Amino acid response by Halofuginone in Cancer cells triggers autophagy through proteasome degradation of mTOR

Fig. 6

Halofuginone induces the detachment of mTORC1 from the lysosomes and partial proteasome degradation of mTOR. a WRO cells were exposed or not to 100 nM halofuginone for 8 h and subcellular fractions were separated by discontinuous sucrose gradient from 15 to 55% sucrose density. Nine fractions were collected and characterized by immunoblotting with the indicated antibodies. A strong reduction in the content of mTOR and RAPTOR proteins is clearly appreciable in fraction 4 of WRO cells treated with HF. A similar trend was observed in two other independent experiments and with diverse sucrose gradient. Post-nuclear supernatant (PNS) and gradient loading fraction (L) were loaded as controls. FR.N.: fraction number. b WRO cells were exposed or not to 100 nM of halofuginone (HF), or 10 μM of the proteasome inhibitor MG132, or both. After 8 h, the cells were collected and homogenates processed for immunoblotting to assess mTOR protein level. As loading control, the filter was stripped and probed with anti β-Tubulin antibody. The pattern of protein expression shown was reproduced in three separate experiments. Densitometry of the protein bands was performed and average mTOR/Tubulin ratios are shown in the histogram graph. Error bars: standard deviation. Statistically significant differences between mTOR protein levels are shown (*, p ≤ 0.05)

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