Rhesus Macaque Fibroblast

Proteasome inhibition Campbell et al 2008 Express native levels of untagged TRIM5α. Can see RAVs in fibroblasts for the first time. Most of these cells actually cells displayed mainly auto fluorescent puncta and probably did not correlate to any virus because light image only taken in the red channel. Discovered the autofluorescent problem when looking at fibroblasts with no virus, bright puncta everywhere and correlate to onions ; Tilt Series Date: 2015-05-18 Data Taken By: Stephen Carter Species / Specimen: Rhesus Macaque Fibroblasts infected with Vpr-Ruby HIV-1 Strain: Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 1°, constant angular increment, dosage: 180.0 eV/Ų, defocus: -10.0 μm, magnification: 18000x. Microscope: Caltech Polara Acquisition Software: UCSFTomo Upload Method: reupload_downloaded Processing Software Used: IMOD Collaborators and Roles: Fibroblasts VSV-G pseudotyped virus Vpr-Ruby provided by Tom Hope Purification / Growth Conditions / Treatment: Cells infected with ZLK 106-06-06-14 HIV-1 GFP Vpr-Ruby. 500 µl of media and 500 µl of virus. Virus spinoculated at 16°C at 18 g for 1 hr. Cells were washed with warm media and incubated at 37°C. Cells frozen at early time points approximately 20 mins after temperature shift. Proteasome inhibition 200mg/ml ; Files available via S3 at https://renc.osn.xsede.org/ini210004tommorrell/tomography_archive/scd2015-05-18-8 tomography7.rec, Reconstruction (Pixel Size 24.0 nm), 890.9 MB Download tomography7.mrc, Tilt Series (Pixel Size 6.0 nm), 3.4 GB Download tomography7_full.rec, Reconstruction (Pixel Size 0.0 nm), 533.4 MB Download