FIG. 5.

Knockdown of trans-acting factors stabilizes AANAT 3′UTR-containing mRNA. (A and B) HEK-293T cells were cotransfected with siRNAs (mock, hnR283, hnL1066, and hnR283/hnL1066 siRNA) and with reporter plasmids pcNAT and pcNAT-3′N578. (A) The level and specificity of hnRNP R and hnRNP L depletion at 36 h after transfection was assessed by Northern and immunoblot analyses. (B) AANAT levels were determined by immunoblot (IB) analysis using antibodies against AANAT and GAPDH. Activities and mRNA levels of AANAT in pcNAT-3′N578- or pcNAT-transfected cells were determined as described in the legend to Fig. 4A.