Figure 2

EGFP expression in dissociated adult DRG cultures after acute exposure to lentivectors. Primary cultures of DRG cells dissociated from adult rats were transduced at MOI = 20 using VSV-G pseudotyped lentivectors encoding EGFP driven by UbC, CAG, and EF1α promoters, or using RABPV pseudotyped EF1α-EGFP, all packaged by the second-generation system. Cultures were maintained 5-DIV before colocalization immunofluorescence (IF), cell identification, and quantitative analysis. Representative images of the EGFP-expressing dissociated DRG cultures (A, B, C, and D) and their correspondent β3-tubulin IF images (E, F, G, and H) revealing neuronal patterns, merged data for identification of EGFP-positive neurons (I, J, K, and L), phase-contrast images captured in the same fields (M, N, O and P), and higher magnification images showing neuronal somata and neurite projections in transduced neurons (Q, R, S and T), as well as transduced non-neuronal cells colocalized with glutamine synthetase (U, V, W and X), are shown in the panels of far-left column for VSV-G UbC, the second-left column for VSV-G CAG, the second-right column for VSV-G EF1α, and the far-right column for RABPV EF1α. Filled arrows point to transduced neurons, empty arrows to non-transduced neurons, filled arrowheads to transduced neuronal projections, and empty arrowheads to transduced glia. Scale bars = 100 μm.