Figure 4.

AIB1 regulates polyribosome recruitment to cancer-related targets.

qRT-PCR analysis was performed on total and fractionated mRNA samples following polysomal fractionation obatined from lentiviral stably expressing shRNA scrambled control (shCON) and AIB1 (shAIB1) MCF-7L cells. Primers were used as probes to measure relative changes in gene expression of total RNA and in light (1-6; lower percentage of ribosome-bound mRNA) and heavy (7-10; higher percentage of ribosome-bound mRNA) pooled ribosomal fractions; A) 4E-BP1, B) eIF2α, C) c-MYC, D) BCL2 and E) eIF5. Graphs for total RNA depict three treatment replicates comprised of three technical replicates. Error bars are SEM. Independent examples of a single experiment for each gene are shown of the ribosomal fractions. RPLPO primers were used as a control for each gene examined. The graphs represent the mRNA copy number (arbitrary value) of each reference gene normalized to RPLPO.

F) Immunoblot analysis was performed on protein lysates obatined from shCON and shAIB1 MCF-7L cells. Total MAPK was used as a loading control.