Skip to main content
. 2017 Dec 13;46(3):1457–1469. doi: 10.1093/nar/gkx1237

Figure 6.

Figure 6.

mTOR controls LARP1 repressive function and affinity for TOP sequences. (A) LARP1 repressive activity is increased by mTOR inhibition. LARP1497–1019 was purified from HEK-293T cells treated with vehicle (DMSO) or 250 nM Torin 1 for 2 h. A total of 0.5 ng TOP or non-TOP (nTOP) synthetic mRNAs were then translated in control-treated extracts in the presence of protein storage buffer (control) or 100 nM LARP1497–1019 from control- (LARP1497–1019-Ctl) or Torin 1-treated (LARP1497–1019-T1) cells, as indicated. Reactions were subsequently analyzed by luciferase assay, and values were plotted relative to control reactions with no LARP1497–1019 added. Significance was calculated by t-test relative to control reaction. (B) LARP1 affinity for TOP sequences is increased by mTOR inhibition. LARP1497–1019 was purified from HEK-293T cells treated with vehicle (DMSO) or 250 nM Torin 1 for 2 h as in (A). Binding of 100 nM LARP1497–1019 to capped TOP RNA probes from (A) was analyzed by EMSA. (Top left panel) SYPRO Ruby staining of input protein. (Bottom left panel) Representative gel showing bound and unbound RNA. (Right panel) Quantification of binding. (n = 4, error bars are SD, significance by t-test).