Figure 2. Depletion of endogenous citron-K inhibits HIV-1 virion production.
(A and B) Knockdown of endogenous citron-K using citron-targeted shRNA constructs in 293T cells inhibited HIV-1 virion production. 293T cells were transfected with citron-targeted shRNA or control plasmid constructs, 24 h later, cells were co-transfected with pNL4GFP and citron-targeted shRNA or control plasmid constructs. Supernatant and cells were harvested 48 h after the second transfection and analyzed for cell-associated gene expression of citron-K, p24 and actin (A) and relative HIV-1 virion production in the supernatant (p24 pg/mL) (B). (C and D) Knockdown of endogenous citron-K in T cells inhibited HIV-1 virion production. Jurkat T cells were transfected with pNL4GFP and citron-targeted shRNA or control plasmid constructs. Supernatant and cells were harvested at 48 h post-transfection. The effect of citron-K knockdown on HIV-1 virion production in the supernatant was measured by p24 enzyme-linked immunosorbent assay (C). *P < 0.05. Cells were lysed and analyze with anti-citron and anti-actin antibodies (D).