Figure 3

Expression of hnRNPs in MDM. A: Western blot detection of hnRNPs A1, A2/B1 and H in nuclear extracts from uninfected and HIV-infected MDM from the same donor (donor 1) over a 5-week period following infection. Representative of 6 cultures. B: Quantification by densitometry (Odyssey Infra-red Imaging System, Li-Cor) of hnRNP expression in 3 representative MDM cultures (Donors 1–3) following infection relative to expression in matched uninfected cells at each time point (Exp [I] : Exp [UI]), showing the donor-to-donor variability commonly seem with MDM cultures. Mean expression levels are shown in the bottom right hand panel (mean + SEM of 6 donor cultures). C: Data from B above normalized to expression levels at 1 week post-infection. D: Confocal laser scanning microscopy of hnRNP A1 expression in the nuclei of MDM, from a different donor from those used in A and B above, exposed to virus or mock infected, 1 and 5 weeks after infection. The replication kinetics for this donor are shown in the top right hand panel. Specific nuclear fluorescence (Fn-b) of at least 10 cells per chamber at weekly intervals after infection was determined using Image J software after subtracting the background fluorescence from cells stained without primary antibody (mean + SEM; bottom left hand panel). The nuclear fluorescence of infected cells (Fn-b [I]) relative to uninfected cells (Fn-b [UI]) over the 5 weeks of infection for this culture is also shown (mean + SEM; bottom right hand panel). Representative of 6 donors.