Production of infectious lentiviral particles from transfected 293 T cells. A) 293 T cells were mock transfected, or transfected with pDNL6, pVSV-G, and either pHR'CMV or pHR'CMV:myc-p30, and supernatants were western blotted for HIV p24 Gag expression. B) Lymphocytes co-cultured with 293 T producer cells transfected with either pHR'CMV or pHR'CMV:myc-p30 were harvested, fixed and stained with anti-myc (9E10) and anti-mouse-Alexa488, and visualized by immunofluorescence. C) Lymphocytes co-cultured with 293 T cells transfected with either pHR'CMV or pHR'CMV:GFP were fixed and analyzed for GFP expression by flow cytometry. D) Cytosolic or nuclear protein fractions were isolated following cytoplasmic RNA extraction, and were analyzed by western blotting with either anti-cyclin A or anti-Bax to assess nuclear and cytoplasmic fraction purity, respectively.