Fig. 2. Cystatin B inhibits the nuclear translocation of USTAT-1 in HIV-infected MDM.
Nuclear translocation of STAT-1 was observed in HIV-1 infected MDM treated with cystatin B siRNA. At 12DPI, HIV-infected MDM were assayed by immunofluorescence for cystatin B (green) and USTAT-1 (red). Blue fluorescent DAPI was used as nuclear stain. Non-targeting siRNA was used as siRNA control. Cystatin B was silenced in MDM cultures using siRNA. Uninfected MDM expressing cystatin B showed a predominantly cytoplasmic localization of USTAT-1 (1–3). After cystatin B gene silencing, STAT-1 was retained in the cytoplasm of uninfected MDM (4–6) as well as in HIV infected MDM treated with non- targeting siRNA (7–9). At 12 DPI, USTAT-1 was totally translocated to the nucleus in HIV-infected MDM after cystatin B gene silencing (10–12). Images were obtained on a Nikon Eclipse E400 fluorescence microscope with a SPOT Insight QE camera and SPOT 5.1, 40x. These results are representative of two independent experiments performed using two different donors.
