ASO-mediated increase in mir-1225 corresponds with a decrease in the abundance of predicted target RNAs. (A) Gel of endogenous miR-1225 and Pkd1 stem–loop RT-PCR and RT-PCR products, respectively, amplified from RNA isolated from cultured cells treated with 12.5, 25, 50, 100, 200 nM of ASO-15 or -16. miR-877 is a loading control. (B) Quantitation of results in (A). Half-maximal effective concentration (EC50) was determined after fitting the data using nonlinear regression with a variable slope. (C) RT-PCR analysis of Jak1, Keap1, and Clcn5 cDNA from cultured cells treated with 12.5, 50, or 200 nM of ASO-16. C refers to control, vehicle treatment. (D) Quantitation of amplicons in (C) with each sample normalized to Gapdh and graphed relative to mock-treated sample. Error bars are SEM. n = 3–6 as indicated by individual points. (*) P < 0.05, (**) P < 0.01, (***) P < 0.0001. One-sample t-test relative to control-treated cells.
