Fig. 4.
NRAD1 is regulated by ALDH1A3 and all-trans retinoic acid. a QPCR analysis of expression of NRAD1 and the other Aldefluorhigh-enriched lncRNA in SUM149 cells (n = 7) or MDA-MB-468 cells (n = 4) following knockdown of ALDH1A3 with two separate shRNAs. ND = not detected (levels below quantification threshold). b QPCR analysis of ALDH1A3 and NRAD1 levels following overexpression (OE) of ALDH1A3 in MCF7 cells (n = 4). a, b Transcript levels are shown normalized to reference genes PUM1 and ARF1 and then log2 fold change over control samples. Western blot confirming changes in ALDH1A3 levels is below, using total protein as a loading control (Supplemental Fig. 9 shows uncropped blots). c RNA-seq co-expression of NRAD1 and ALDH1A3 in the TCGA Cell 2015 dataset (all breast cancer patients) was retrieved using cBioportal. d RNA-seq co-expression of NRAD1 and ALDH1A3 in the Cancer Cell Line Encyclopedia (only breast cancer cell lines) was retrieved using the CCLE portal. e QPCR analysis of NRAD1 levels in a panel of cell lines treated with 100 nM retinoic acid for 24 h versus no treatment (n = 4). Significance was determined using a one-way ANOVA with Dunnett’s post-test (a, e), or a student’s t-test (b). c, d, Spearman and Pearson correlations are shown, and significance was determined using linear regression. All error bars represent standard deviation