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. 2016 Jun 30;7(31):48887–48904. doi: 10.18632/oncotarget.10346

Figure 4. MYC is required for the response of rDNA transcription to amino acids.

Figure 4

A. HeLa cells were transfected with either non-targeting siRNA control (siCON) or pooled siRNA against MYC (siMYC) for 3 days, amino acid and serum-starved for 2 hours and then stimulated with amino acids (AA) for 3 hours. Cells were pulse labeled and 47/45S rRNA synthesis analyzed. Representative images and resultant graph (mean +/− SEM) of n = 3 experiments. Below the graph are representative immunoblotting images, n = 2. * p < 0.05 compared to Lane 2; # p < 0.05 compered to lane 4. B.-D. HeLa cells were amino acid and serum starved (-AAS) for 2 hours and then stimulated with all amino acids (AA) for the times indicated B. and D. or 3 hours C. Representative immunoblotting images B., n = 2 blots) and quantitation of qRT-PCR for MYC and MYC transcriptional targets TERT, CCND1, p21 and GAS C. and D., n = 3-4 experiments). *p < 0.05, **p < 0.01 compared to -AAS cells.E. HeLa cells were deprived of amino acids in the presence of 10 μM MG132 for 3 or 6 hours. Representative immunoblotting images from n = 2 experiments. F. HeLa cells were amino acid and serum-starved for 2 hours and then stimulated with amino acids in the presence of 10 μg/ml cyclohexamide (CHX) for the times indicated. Representative immunoblotting images and quantitation (mean +/− SEM) from n = 3 experiments. G. HeLa cells were amino acid and serum-starved for 2 hours, pre-treated with 20 nM rapamycin for 30 minutes prior to 3-hour amino acid stimulation. Representative of n = 2 blots.