Figure 5.

Representative siRNA-directed inhibition of HIV-1 replication. (A) Using two candidate siRNAs per gene shown, each siRNA was transfected into HIV-1 latently infected OM-10.1 cells at mid log phase of growth. Following transfection, viral activation, and treatment, supernatants were collected and analyzed for p24 Gag expression by ELISA. The white crossed bars represent the first set of experiments, while the black bars represent the second run performed in an identical manner. (B) For Western blots, protein samples (one hundred micrograms of each extract) were separated on SDS-PAGE and then transferred to an Immobilon-P (polyvinylidene difluoride; Millipore) membrane and blocked with 5% fat-free milk (in TNE50/0.1% Nonidet P-40). Membranes were incubated overnight with various primary antibodies, and reactive complexes were developed with protein G-labeled ¹²⁵I and visualized with a PhosphorImager scanner (Amersham Biosciences).