![Figure 2. Hsp70 specifically inhibits nuclear import of Vpr. (A) HEK 293T cells were transfected with vectors expressing indicated constructs.27 At 48 hours after transfection, cells were fixed, mounted in ProLong Antifade Imaging Medium (Molecular Probes, Eugene, OR), and analyzed by fluorescent microscopy. (B) HEK 293T cells were cotransfected with an Hsp70-expressing (or an empty pcDNA) vector and a plasmid encoding indicated constructs. At 48 hours after transfection, cells were exposed to leptomycin B (LMB, 400 nM) for 2.5 hours, fixed, and analyzed by fluorescent microscopy. A 5-fold excess of the Hsp70-encoding vector over the plasmid-encoding Vpr construct was used. All images were acquired using a BX-60 Olympus Epi-fluorescence microscope (Melville, NY) with Evolution MP Digital Camera using Image-Pro Plus v. 4.5 acquisition software (Media Cybernetics, Silver Spring, MD). Images are presented at × 100 original magnification (100 × oil objective lens [NA = 1.30]). Results are shown for 1 representative experiment of 3 performed.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/104/6/10.1182_blood-2004-01-0081/6/zh80180466790002.jpeg?Expires=1738086491&Signature=zTrZm0g1lVWs2CQCgoIEJf3f-z0RY8ILAKocQlBG7Tpki8nrQTtgdTMvmYX8zm4nekoul7gPEsHS89BlxSX8yL1sTuE~PkE6jdnhPNc~xekxW3nRSSoZoFBqG8Rmilbi~0aFFbRowE7S1IaEa0u5vlqL2ZadtYlmrKWizsCZDdZv6-tqKQ4MKmWNNJPqFLcyd6vQ2ApmTfG1AeY~lnMs5qv0x-0rGYoXS1Fw5TOQKfA6sq85dUeCA1hfhKQDTmXy7CczYwZf04KPDw3pyDkP1RcYH7PU8Rp6Q~7htAFsLVKTIv4AMTa1lDnWWT-UqFEQOqgq9Q8U7bY0X1mMxJ9PVg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Hsp70 specifically inhibits nuclear import of Vpr. (A) HEK 293T cells were transfected with vectors expressing indicated constructs.27 At 48 hours after transfection, cells were fixed, mounted in ProLong Antifade Imaging Medium (Molecular Probes, Eugene, OR), and analyzed by fluorescent microscopy. (B) HEK 293T cells were cotransfected with an Hsp70-expressing (or an empty pcDNA) vector and a plasmid encoding indicated constructs. At 48 hours after transfection, cells were exposed to leptomycin B (LMB, 400 nM) for 2.5 hours, fixed, and analyzed by fluorescent microscopy. A 5-fold excess of the Hsp70-encoding vector over the plasmid-encoding Vpr construct was used. All images were acquired using a BX-60 Olympus Epi-fluorescence microscope (Melville, NY) with Evolution MP Digital Camera using Image-Pro Plus v. 4.5 acquisition software (Media Cybernetics, Silver Spring, MD). Images are presented at × 100 original magnification (100 × oil objective lens [NA = 1.30]). Results are shown for 1 representative experiment of 3 performed.
