Figure 2
From: Characterization of a potent non-cytotoxic shRNA directed to the HIV-1 co-receptor CCR5
The decreased levels of CCR5 expression by CCR5 shRNA (1005) correlates with that of CCR5 mRNA. huCCR5-293T cells were transduced with lentiviral vectors bearing either shRNA 1005 against CCR5 {CCR5shRNA (1005)} or a control shRNA against firefly luciferase (Luc shRNA). To monitor the expression levels of CCR5 on cell surface, the cells were cultured for 4 days and stained with either PE-Cy5 conjugated anti human CCR5 monoclonal antibody or isotype control. CCR5 and EGFP expression were analyzed by flow cytometry. The percentage number in each quadrant is indicated in each panel (A). To measure the levels of CCR5 mRNA, total RNA was isolated using Qiagen RNeasy extraction kit. Quantitative RT-PCR was performed using IQ5 with iScript one step RT-PCR kit using β-actin as an internal control (B).