American Society of Hematology

Figure 6

$Figure 6. Rapamycin reduces c-MYC-ER mRNA translation in differentiating MPRO c-MYC-ER granulocytes. (A) Polysome profiles generated from lysates of AGN194204 (RXR)–treated MPRO c-MYC-ER cells treated with 200 nM 4-hydroxytamoxifen (RXR + 4OHT) or 200 nM 4OHT and 80 nM rapamycin (RXR + 4OHT + rapa) for 4 days and normalized by loading equal cell number. (B) Total RNA per cell was measured using a NanoDrop ND-1000 UV-Vis Spectrophotometer (NanoDrop Technologies, Wilmington, DE). (C) Abundance of c-MYC-ER mRNA per fraction determined by qRT-PCR in c-MYC-ER–expressing MPRO cells treated as described in panel A. Values are normalized to c-MYC-ER mRNA abundance in fraction 1 for MPRO c-MYC-ER cells treated with 4OHT. (D,E) Distribution of Eef1α, β-actin, and c-MYC-ER mRNA in fractionated lysates was determined by qRT-PCR. For each individual transcript, results are expressed as the percentage of the sum of the transcript mRNA present in 12 fractions. Data in panels A through E are from a single representative experiment.$

Rapamycin reduces c-MYC-ER mRNA translation in differentiating MPRO c-MYC-ER granulocytes. (A) Polysome profiles generated from lysates of AGN194204 (RXR)–treated MPRO c-MYC-ER cells treated with 200 nM 4-hydroxytamoxifen (RXR + 4OHT) or 200 nM 4OHT and 80 nM rapamycin (RXR + 4OHT + rapa) for 4 days and normalized by loading equal cell number. (B) Total RNA per cell was measured using a NanoDrop ND-1000 UV-Vis Spectrophotometer (NanoDrop Technologies, Wilmington, DE). (C) Abundance of c-MYC-ER mRNA per fraction determined by qRT-PCR in c-MYC-ER–expressing MPRO cells treated as described in panel A. Values are normalized to c-MYC-ER mRNA abundance in fraction 1 for MPRO c-MYC-ER cells treated with 4OHT. (D,E) Distribution of Eef1α, β-actin, and c-MYC-ER mRNA in fractionated lysates was determined by qRT-PCR. For each individual transcript, results are expressed as the percentage of the sum of the transcript mRNA present in 12 fractions. Data in panels A through E are from a single representative experiment.

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