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. 2019 Jan 29;10(2):96. doi: 10.3390/genes10020096

Figure 2.

Figure 2

Translation of endogenous β-actin mRNA decreases under amino acid starvation and mTOR inhibition. (A–B) HEK293T cells were treated for 2h with DMSO (control), 500 nM Torin1 (A), or subjected to amino acid and serum starvation (B). The cells were lysed, fractionated at a sucrose gradient of 15–45%, and mRNA abundance of endogenous SLU7, ACTB, RPL32, RPL14 and YB-1 in polysome- and subpolysome fractions was measured by qPCR. A polysome fraction was estimated as (mRNA in polysome)/(mRNA in polysome + mRNA in subpolysome). (C–D) HEK293T cells were transfected with pNL2.2; after 2h of amino acid and serum starvation (-AA) the cells were collected, and luciferase activity (C) and NlucP mRNA abundance (D) were measured. Student’s t-test was used to estimate the statistical significance versus control treatment (A–B) or versus the SLU7 control (C–D), *** p < 0.001; *p < 0.05; #—nonsignificant.