Identification of cellular deoxyhypusine synthase as a novel target for antiretroviral therapy
J. Clin. Invest. Ilona Hauber, et al. 115:76 doi:10.1172/JCI21949 [Go to this article.]

Figure 5
Inhibition of DHS by RNAi. For DHS knock-down, HLCD4-CAT cells, which express the CD4 surface molecule, were repeatedly transfected with either human DHS–specific (DHS) or control scramble II duplex (SDII) siRNAs. (A) RNA was isolated 1 day after the final transfection, followed by first-strand cDNA synthesis. Gene silencing was monitored by DHS-specific PCR. A GAPDH-specific reaction served as internal control. (B) siRNA-treated cells were metabolically labeled using [¹⁴C]spermidine, and total cell proteins were analyzed by 2-dimensional electrophoresis and subsequent autoradiography as described in the legend to Figure 1. (C and D) The respective cell cultures were infected with HIV-1 NL4/3 1 day after the final transfection. At day 3 after infection, p24 antigen levels (C, black bars), cell viability (MTT assay; C, gray bars), and integrated proviral DNA (D) were analyzed. NC, negative control reaction.