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. 2004 Apr 30;101(19):7347–7351. doi: 10.1073/pnas.0402107101

Fig. 2.

Fig. 2.

(A) 293T cells expressing two copies of GFP were infected with different lentiviral constructs, with mois as shown in B. Vector titers were determined by p24 ELISA and by TaqMan real-time PCR determination of transduced proviral genomes. On average, the vector preparations were 1.5 E4 particles per nanogram of p24. Cells were cultured for 9 days, and GFP expression levels were quantitated by FACS analysis. GFP expression levels were unaffected by infection with LV-CRE alone (Aa). Infection with LV-shGFP-OFF in the absence of LV-CRE resulted in no change in GFP levels (Ab), whereas LV-shGFP-WT and LV-shGFP-ON were equally effective in down-regulating the target (A c and d). Coinfection of LV-CRE (moi = 70) and LV-shGFP-OFF (moi = 100) resulted in efficient down-regulation of GFP expression (Ae). (B) FACS analysis composite. Coinfection of LV-CRE (moi = 70) with decreasing mois of LV-siGFP-OFF (moi = 60, 20, or 10) resulted in decreased silencing of GFP (data not shown). Similarly, decreasing of LV-CRE (moi = 70, 35, 7, and 3) while keeping LV-shGFP-OFF constant (moi = 100) diminished silencing (data not shown).