Fig. 6

5’TOP motif reporter is less sensitive to starvation in aneuploid cells. a Schematic illustration of the pS-16-wt-luc luciferase reporter construct. RPS16 transcriptional start side and 29 nt of Exon 1 including the 5’TOP motif is incorporated into the 5’UTR of luciferase gene. b pS16-wt-luc activity after overexpression of hsa-miR-10a-5p. Cells were reverse transfected with hsa-miR-10a-5p mimic or control molecule and 24 h post transfection forward transfected with pS16-wt-luc and pRL-TK. 72 h post mimic transfection the luciferase assay was conducted. c pS-16-wt-luc activity after starvation. Cells were transfected as in B and serum- starved for 3 h before the luciferase reporter assay was conducted. Data presents Renilla normalized mean +/− SD values. Two-way ANOVA, multiple comparison correction with Sidak test. ns = not significant, *P < 0.0332, **P < 0.0021, ***P < 0.0002, ****P < 0.0001. d Abundance of has-miR-10a-5p determined by quantitative real-time PCR upon prolonged cyclohexamide treatment. e Endogenous hsa-miR-10a-5p activity upon prolonged cyclohexamide treatment determined by psiCheck2-10a luciferase reporter assay with hsa-miR-10a-5p binding site