Type I Interferons and Interferon Regulatory Factors Regulate TNF-Related Apoptosis-Inducing Ligand (TRAIL) in HIV-1-Infected Macrophages
Figure 4
siRNA knockdown of IRF-1 and IRF-7 reduces STAT1 phosphorylation and TRAIL expression in HIV-1-infected macrophages.
Two days after HIV-1 infection, MDM were transfected with siRNA for IRF-1, -3, or -7. A. Forty-eight hours later, successful transfections were confirmed by Silencer FAM-labeled Negative Control #1 siRNA transfection indicator (green). Hoechst 33258 (nucleus marker, blue) was used to visualize the total cell number. B–D. Total RNA was collected 48 hours post-transfection and mRNA levels of IRF-1(B), -3(C), or -7(D) were determined by real-time RT-PCR. E. Ninety-six hours after transfection, p-STAT1 and total STAT1 were detected by Western blotting. β-actin was used as a loading control. F. TRAIL expression levels were determined by real-time RT-PCR. Results were normalized with GAPDH and shown as the fold change over non-specific siRNA control. G. Supernatants were tested for HIV-1 RTase activity. ** indicates p<0.01 when compared to control; # indicates p<0.05 when compared to HIV group with siRNA control; ## indicates p<0.01 when compared to HIV group with siRNA control. Data are representative of three donors.