Unravelling the RNA-Binding Properties of SAFB Proteins in Breast Cancer Cells

Figure 4

SAFB2 regulates expression of MALAT-1. (a) Expression of MALAT-1 was measured by qRT-PCR using RNA from MCF-7 and MDA-MB-231 cells transfected with negative, SAFB1, SAFB2 or SAFB1 and SAFB2 siRNA using validated TaqMan probes specifically targeting MALAT-1. Data represents the average of three biological replicates ± SD. Repeated Measures ANOVA with Dunnett’s Multiple Comparison Test statistical significance of mRNA expression was calculated using Student’s -test; . Enrichment of MALAT1 RNA by conventional PCR (b) and qPCR (c) after RNA immunoprecipitation with anti-SAFB1 in MCF-7 cells. No enrichment was observed using IgG. (d) Intranuclear distribution of SAFB1 and SRSF2 in MCF-7 cells by immunofluorescent staining. Confocal laser microscopy revealed a punctate pattern for SAFB1 and SRSF2 in nuclear speckles.

(a)

(b)

(c)

(d)