Figure 1.

HuR associates with the endogenous DNMT3b mRNA. (A) The four alternatively spliced transcript variants for the DNMT3b gene. (B) DNMT3b 3′UTR showing one predicted HuR motif hit. (C) Two days after siRNA transfection, RKO cells were harvested for western blot analysis to monitor the expression of HuR and loading control α-Tubulin. (D) RKO cells were transfected with either control (C) or HuR siRNA and the lysates were used in IP reactions employing anti-HuR antibodies or control IgG1; RNA was subsequently isolated and used in RT reactions (inset). Representative PCR products of all DNMT3b variants visualized in ethidium bromide-stained agarose gels; the levels of GAPDH (a housekeeping mRNA which is not a target of HuR) served to verify equal sample input. (Graph) Fold differences in DNMT3b variant 3 mRNA abundance in HuR IP compared with IgG IP, as measured by RT–qPCR analysis. The means and standard error of the means (SEM) from three independent experiments are represented (**P ≤ 0.01).