Figure 4.

HOTAIR is transcriptionally regulated by FOX proteins and repressed by oestrogen. (A) Top, western blot for indicated proteins, demonstrated siRNA knockdown in MCF7 cells and below qRT-PCR results for HOTAIR expression in each knockdown, normalized to RPLP0 and the scrambled control. (B) Expression of HOTAIR and FOXM1 following treatment of MCF7 cells by DMSO (vehicle) or thiostreptin (ThSt) (C) Re-analysis of MCF7 GRO-Seq data following E2 treatment [GSE27463] (81). The yellow histograms represent the positive strand and blue, the negative. The X-axis corresponds to the genomic position from Hg18 and the Y-axis the relative reads, HOTAIR promoters are shown above gene track. (D) RNA-Seq results for HOTAIR from E2 treated MCF7 cells over 72 hours (Reads per kilobase per million = RPKM). (E) qRT-PCR results of HOTAIR expression from a 10 nM E2 time course in MCF7 cells, representative of three separate experiments, data not shown. Y-axis represents relative expression (ΔΔCt of RPLP0/DMSO). Statistical analysis was performed via a one-way ANOVAs with Tukey’s corrected multiple comparisons between indicated columns or data points for A and E and via a two-tailed t-Test for B, P-values are <0.05 (*), <0.01 (**), <0.001 (***) and <0.0001 (****).