Figure 2
Effect of REST on REST-003 ncRNA expression in cells and effect of si-REST-003 on MDA-MB-231 invasiveness.
(A) (B) Effect of REST on expression of REST-003 ncRNAs in si-REST-treated MCF-7 (A) and REST-overexpressed MDA-MB-231 cells (B) by qRT-PCR. MCF-7 cells treated with a non-REST siRNA (si-GAPDH) (A) and MDA-MB-231 cells transfected with EGFP or mt-REST cDNA6 (lacking two repressor domains) (B) served as controls. ‘REST Rel. Exp.’ refers to REST‐001 (primer set R‐N). (C) Detection of REST-003 ncRNA by qRT-PCR (left) and invasiveness by a Matrigel invasion chamber (right) after treating MDA-MB-231 cells with si-REST-003. (D) Reduced REST-003 expression by si-REST-003 treatment in MDA-MB-231 cells using qRT-PCR. More than 50% of REST-003 transcripts were reduced by si-REST-003 relative to si-C (scramble). REST transcript expression [REST-001 (R-001), R-M primer pair] was not changed by si-REST-003 (si-R-003) treatment. For qRT-PCR data, expression levels were normalized to GAPDH, CyclophilinA and/or Actin, converted to MNE and presented as Rel. Exp. after normalization to control cells (A, B) or si-C samples (D). Biological replicates are shown on the bar graph as mean plus SEM [one-way ANOVA with Friedman test for multiple comparisons for (A, B) and paired t-test for (D)].
