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. 2018 Feb 7;9:63. doi: 10.3389/fphar.2018.00063

FIGURE 3.

FIGURE 3

Substrate-induced regulation of SNAT2 stability. HeLa cells were transiently transfected with 10 μg of a pcDNA6 vector containing (A) SNAT2V5, (B) SNAT5V5, or (C) SNAT2-5V5; 48 h post-transfection, cells were incubated for 8 h in the absence or presence of AAs or in EBSS containing 2 mM of the indicated AA. Lysates (40 μg) were then subject to SDS–PAGE and immunoblotting with an anti-V5 antibody or antibody directed against actin, which was used as a gel loading control. Blots are representative of three separate experiments, bar graphs values are mean ± SEM. V5 expression was quantified using ImageJ software and results expressed as a fold change in expression of the transporter relative to that in cells incubated in the presence of AAs. Asterisks indicate a significant difference between the indicated bars (P ≤ 0.05).