Figure 5.
Dominant-negative effects on endo-siRNA biogenesis by overexpression of Loqs isoforms. (A) Detection of endogenous Loqs protein from S2-cell extract after immunoprecipitation with α-Dcr-1, α-Loqs-PB C-terminus and α-Loqs-PD; rb IgG and α-R2D2 served as controls; α-Loqs monoclonal antibody was used for detection of endogenous Loqs isoforms. (B) Northern blot from Drosophila S2 cell extract overexpressing Loqs isoforms; transfection with pUC18 served as a control; L1L2 = Loqs truncation lacking the PD-specific amino acid sequence, L1L2:L3PC = reconstituted Loqs-PC; myc-loqs-PD (genomic) = expression construct derived from genomic DNA; DNA probes against the long hairpin-derived endo-siRNA CG4068B (66) and against bantam miRNA were used; 2S rRNA served as a control for loading; the two parts were extracted from a single blot using the same exposure and image processing settings.