GAS5 suppresses c-Myc expression at protein level.

<p>(<b>A</b>) GAS5 was knocked down by siRNA in HEK-293T cells, and global protein translation was measured by [³H]-leucine incorporation assay. Bars represent mean ±SD from 3 replicates for Q-PCR and 6 replicates for [³H]-leucine incorporation assay. The experiments were performed two times. (<b>B</b>) pRF and pRF-SL plasmid were transfected into HEK-293T cells and cap-dependent protein translation efficiency was evaluated by luciferase assay. Bars represent mean ±SD from 4 replicates. The data were repeated three times. (<b>C</b>) The protein level of c-Myc (P = 0.007), Mcl1 (P = 0.88), survivin (P = 0.47) and Bcl2 (P = 0.47) was assessed by western blot (n = 3) after the cells were treated with GAS5 or control siRNA. (<b>D</b>) The mRNA level of c-Myc (P = 0.10), Mcl1 (P = 0.75), survivin (P = 0.31) and Bcl2 (P = 0.19) was quantified by Q-PCR after the cells were treated with GAS5 or control siRNA. Bars represent mean ±SD from 3 replicates. The experiment was repeated three times. (<b>E</b>) The protein level of c-Myc (P = 0.005) was assessed by western blot (n = 3) after the cells were transfected with <i>in vitro</i> transcribed GAS5 RNA. (<b>F-G</b>) The mRNA level of c-Myc (P = 0.37) (<b>F</b>) and GAS5 (<b>G</b>) was quantified by Q-PCR after the cells were transfected with <i>in vitro</i> transcribed GAS5 RNA. Bars represent mean ±SD from 3 replicates. The experiment was performed three times.