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Figure 5 | BMC Veterinary Research

Figure 5

From: Constitutive phosphorylation of the mTORC2/Akt/4E-BP1 pathway in newly derived canine hemangiosarcoma cell lines

Figure 5

Effects of serum stimulation on the MAPK/Erk and AKT/mTOR pathways. ( A) Western blot analysis for the expression of Erk, Akt, mTOR, p70S6K, and 4E-BP1. Canine HSA cell lines were grown in Medium 199 with 10% FBS. After 24 h-serum starvation, the medium was replaced with Medium 199 containing 1% FBS or Medium 199 containing 10% FBS for 30 min. The total protein was then extracted using RIPA Lysis Buffer, and equal amounts of protein (10 μg) were subjected to western blot analysis. β-actin was used as a loading control. ( B) Western blot analysis for the expression of PTEN. The total protein was extracted from subconfluent cells grown in Medium 199 with 10% FBS using RIPA Lysis Buffer, and equal amounts of protein (10 μg) were subjected to western blot analysis. β-actin was used as a loading control.

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