FIG. 5.

RNAi of ALF in Hpt cell cultures detected by in situ hybridization. One microgram of ALF or GFP dsRNA/well was transfected into the Hpt cell cultures on coverslips using histone H2A as transfection reagent. In situ hybridization was performed 3 days post-dsRNA transfection using a digoxigenin-labeled ALF cDNA fragment as a probe. Hpt cells without probe or RNase H-treated Hpt cells were also used as negative controls. The fluorescent signal was detected by confocal microscopy. (A) ALF dsRNA-silenced cells; (B) GFP dsRNA-treated cells; (C) control cells without any probe; (D) RNase H treatment of control cells. Bars, 10 μm.