Knockdown efficiency and individual repeats of luciferase-PTEN 3’UTR reporter assay in DU145 cells co-transfected with siRNA against PTEN ceRNAs.
This is the same experiment as Figure 1. (A) Representative microscopy images (10X magnification) of DY-547-labeled siGLO RISC-Free control transfected DU145 cells 48 hr after transfection. Transfection efficiency was estimated to be >90%. (B) Independent biological repeats of luciferase reporter assay. Relative luminescence unit (RLU) is presented for each condition relative to the siNC condition. Means reported and error bars represent SD from three technical replicates. (C) Independent biological repeats of RT-qPCR analysis. Expression of each transcript after transfection of its respective siRNA relative to negative control transfection (siNC) is presented. Transcripts listed on y-axis. Means reported and error bars represent SD from three technical replicates. One-sample t-tests of transcript expression data after transfection of respective siRNA to a constant of 1 (relative value of siNC). SERINC1: t(3) = 45.8, uncorrected p=2.30×10−5, Bonferroni corrected p=1.15×10−4; ZNF460: t(3) = 2.98, uncorrected p=0.059, Bonferroni corrected p=0.294; VAPA: t(3) = 8.94, uncorrected p=0.0030, Bonferroni corrected p=0.015; CNOT6L: t(3) = 11.3, uncorrected p=0.0015, Bonferroni corrected p=0.0074; PTEN: t(3) = 11.2, uncorrected p=0.0015, Bonferroni corrected p=0.0077. Additional details for this experiment can be found at https://osf.io/spv4f/.