Fig. 1
From: Suppression of PDHX by microRNA-27b deregulates cell metabolism and promotes growth in breast cancer
In silico and experimental evidence predict the 3’UTR of PDHX is targeted by MicroRNA-27b. a Diagram illustrating the stepwise approach used to select potential miR-27b targets for experimental validation. b Preliminary luciferase assay of MCF7 cells transiently cotransfected with a PDHX luciferase vector and miR-27b mimic or control. Luciferase expression is quantitated by measuring the degree of luminescence detected by a luminometer 24 h post-transfection for each assay. c and d TaqMan qRT-PCR assessing miR-27b levels in transfected MCF7 and 4175 cells to confirm adequate delivery of RNA oligonucleotides into the cell. e Nucleotide sequences indicating the location of miR-27b binding within the PDHX 3’UTR. The sequence of the original luciferase construct and the eight base pair substitutions present in the mutant construct are shown beneath. Luciferase assays in MCF7 (f) and 4175 (g) cells, showing miR-27b targeting of the PDHX-3’UTR sequence is lost upon mutation of the seed region. h Expression of PDHX in the Staunton Cell line dataset accessed online using Oncomine. PDHX Fold Change (vs average of all cancers): − 5.602; p-value: 0.023. Under-expression Gene Rank for PDHX: 335 (in top 7%)