FIGURE 1:

Treatment of HeLa cells with Gle1-SD peptide leads to nuclear accumulation of specific mRNAs. (A) Heat map from RNA-seq data illustrating differences in nuclear RNA accumulated in HeLa cells when treated with 5 µM of Gle1-scr or Gle1-SD peptide. Heat map depicts z-transformed gene expression of all genes that showed a fold change of ≥2 and false discovery rate of ≤0.05. (B) Twelve gene targets from RNA-seq data were confirmed with RT-qPCR in CDS region. The qPCR reads were normalized to actin reads in the respective samples (ΔCT value). The difference between ΔCT values of treated and untreated samples (ΔΔCT) was used to calculate log2 fold change, plotted in the graph. The graph represents fold change values (mean ± SEM) from three biological replicates performed in triplicate. (C) Top panel depicts position of the primers designed to amplify CDS and 3′-UTR (defined in Supplemental Table S2). RT-qPCR analysis of fold change (mean ± SEM) in 3′-UTR of Fos from three biological replicates was performed in triplicate. Samples were normalized to actin (intron 1, In1) using ΔΔCT method. (D) Top panel depicts position of the primers designed to amplify CDS and 3′-UTR. RT-qPCR analysis of fold change (mean ± SEM) in 3′-UTR of FosB from three biological replicates was performed in triplicate. Samples were normalized as in C. ΔCT values were used to calculate statistical significance using one-tailed, paired t test (**p < 0.005, *p < 0.05).