Figure 7

HIV-1 miRNA down-regulated the expression of proteins related to apoptosis. (A) 293T cells were mock transfected (lane 1) or transfected with TAR-D mutant (lane 2) or TAR-WT RNA (lane 3). After 48 hours cell lysates were prepared and Western blotted for ERCC1, PIASγ, GIT2, IER3 or β-actin. (B) Cell lysates were prepared from HeLaT4, HLM-1, CEM, ACH2, U1 and U937 cell lines and Western blotted for ERCC1, PIASγ, GIT2, IER3 or β-actin. Densitometry was performed, and the expression levels were normalized to actin. The average expression level of each protein from three experiments was determined and displayed as the ratio of expression in the infected cells (HLM-1, ACH2 and U937) to their uninfected counterpart (HeLaT4, CEM, U1). (C) HLM-1 cells were transfected with mock (lane 1) or TAR 5' antagomir (lane 2). Cells were lysed after 96 hours and 20 micrograms of protein were used to Western blot for the expression of ERCC1. Coomassie staining of the ~25–50 kDa portion of the gel is included as a loading control.