Fig. 5.

pa-Eleanor(S) plays a role in Eleanor topologically associating domain (TAD) formation and long-term estrogen deprivation (LTED) cell proliferation. a Quantitative reverse transcription polymerase chain reaction showed that pa-Eleanor(S) was knocked down with Antisense LNA GapmeR (LNA) in LTED cells. b Expression levels of genes in Eleanor TAD decreased upon pa-Eleanor(S) knockdown in LTED cells. Values of LNA for the negative control (LNA NC) were set to 1. c RNA fluorescence in situ hybridization (FISH) scanning analysis along the Eleanor TAD, with pa-Eleanor(S) knockdown in LTED cells. BAC clones used as FISH probes are indicated on the panels. The genomic positions covered by the BAC clones are shown in Fig. 1b (green bars). Nuclear Eleanor RNA foci were diminished with pa-Eleanor(S) knockdown. Scale bar, 10 µm. d Quantitative analysis of RNA FISH in c. Total signal intensities per nucleus in LTED cells were measured (n > 40 nuclei per sample). Boxplots shown are center line, median; box limits, upper and lower quartiles; whiskers, 1.5× interquartile range; points, outliers. P values were calculated using two-tailed, Mann–Whitney U test. e Inhibition of LTED cell proliferation due to pa-Eleanor(S) knockdown. Time course analysis was performed after treatment of LTED (left) and MCF7 (right) cells with LNA. Data presented in a, b, e are representative of three independent experiments (mean ± s.e.m.). P values were calculated using unpaired, two-tailed, Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001)