Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Alexa fluor conjugated secondary antibodies |
Thermo Fisher Scientific |
IF (1:1000) | |
| Antibody | Goat anti-mouse IgG Abberior STAR RED |
Sigma-Aldrich; Cat# 52283 |
IF (1:200) | |
| Antibody | Goat anti-rabbit IgG Atto 594 | Sigma-Aldrich; Cat# 77671 |
IF (1:200) | |
| Antibody | Goat polyclonal HRP anti-rabbit IgG | Jackson ImmunoResearch Labs; Cat# 111-035-144 | RRID:AB_2307391 | ELISA (1:2000) |
| Antibody | Mouse monoclonal anti-hCPSF5 | Sigma-Aldrich; Cat# SAB1404890 | RRID:AB_10739397 | IF(1:100) |
| Antibody | Mouse monoclonal anti-hFG repeats (Nuclear Pore Complex) | Abcam; Cat# ab24609 | RRID:AB_448181 | IF(1:300) |
| Antibody | Mouse monoclonal anti-hLamin A/C | Santa Cruz Biotechnology; Cat# sc-7292 |
RRID:AB_627875 | IF (1:100) |
| Antibody | Mouse monoclonal anti-hLEDGF | BD Biosciences; Cat# 611714 |
RRID:AB_399192 | IF(1:100) |
| Antibody | Mouse monoclonal anti-hNup153 | Abcam; Cat# ab24700 |
RRID:AB_2154467 | IF (1:250) |
| Antibody | Rabbit polyclonal anti-hCPSF6 | Atlas Antibodies; Cat# HPA039973 |
RRID:AB_10795242 | IF(1:250) |
| Antibody | Rabbit polyclonal anti-hCPSF7 | Atlas Antibodies Cat# HPA041094 |
RRID:AB_10794187 | IF(1:200) |
| Antibody | Rabbit polyclonal anti-HIV-1 CA | In-house | IF(1:1000) | |
| Antibody | Sheep polyclonal anti-HIV-1 CA | In-house | IF (1:1200) | |
| Cell line (H. sapiens) |
HeLaP4 TNPO3-KD | Z Debyser (University of Leuven, Belgium); Thys et al. (2011) |
||
| Cell line (H. sapiens) |
TZM-bI indicator cells | Wei et al. (2002) | RRID: CVCL_B478 | |
| Cell line (Homo sapiens) |
Human embryonic kidney 293 T cells (HEK293T) | Pear et al. (1993) | RRID: CVCL_0063 | |
| Chemical compound, drug |
Efavirenz (EFV) | AIDS Research and Reference Reagent Program, Division AIDS, NIAID |
||
| Chemical compound, drug |
Flavopiridol | Sigma-Aldrich; Cat#F3055 | ||
| Chemical compound, drug |
iQ Supermix | BioRad; Cat#1708860 | ||
| Chemical compound, drug |
Maraviroc (MVC) | Sigma-Aldrich; Cat#PZ0002 | ||
| Chemical compound, drug |
PF-3450074 (PF74) | Sigma-Aldrich; Cat#SML0835 |
||
| Chemical compound, drug |
Raltegravir (RAL) | AIDS Research and Reference Reagent Program, Division AIDS, NIAID |
||
| Commercial assay or kit |
Click-iT EdU Alexa Fluor 647 Imaging kit |
Thermo Fisher Scientific; Cat#C10340 | ||
| Commercial assay or kit |
DIG-Nick translation mix | Roche; Cat#11745816910 | ||
| Commercial assay or kit |
InviTrap Spin Universal RNA Mini kit |
Stratec; Cat#1060100300 | ||
| Commercial assay or kit |
SuperScript III Reverse transcriptase kit |
Thermo Fisher Scientific; Cat#18080093 |
||
| Commercial assay or kit |
TSA Plus system |
Perkin Elmer; Cat#NEL749A001KT |
||
| Recombinant DNA reagent |
AAV helper plasmid |
D Grimm (University of Heidelberg, Germany) |
AAV helper plasmid expressing rep and cap genes for transducing MDM |
|
| Recombinant DNA reagent |
CPSF6 triple shRNA |
This study | Plasmid expressing three shRNAs targeting CPSF6. For packaging into AAV vector. |
|
| Recombinant DNA reagent |
NS control AAV | K Boerner (University of Heidelberg, Germany); Börner et al. (2010) |
Plasmid expressing non-targeted shRNA. For packaging into AAV vector. | |
| Recombinant DNA reagent |
pEnv-4059 | R Swanstrom (University of North Carolina, USA); Schnell et al. (2011) |
Plasmid expressing an R5-tropic HIV-1 Env | |
| Recombinant DNA reagent |
pHIVSIREN | G Towers (University College London, UK); Rasaiyaah et al. (2013) |
Plasmids expressing shRNAs targeting CPSF6 or non-targeting shRNA | |
| Recombinant DNA reagent |
pMD2.G | D Trono (EPFL, Lausanne, Switzerland) |
RRID: Addgene_12259 | Plasmid expressing VSV-G |
| Recombinant DNA reagent |
pNL4-3 | Adachi et al. (1986) | HIV-1 proviral plasmid |
|
| Recombinant DNA reagent |
pNL4-3ΔEnv | B Müller (University of Heidelberg, Germany) |
HIV-1 proviral plasmid |
|
| Recombinant DNA reagent |
pNL4-3ΔEnv N74D/A77V | This study | HIV-1 proviral plasmid |
|
| Recombinant DNA reagent |
pNLC4-3ΔTat | T Müller (University of Heidelberg, Germany) |
HIV-1 proviral plasmid |
|
| Recombinant DNA reagent |
psPAX2 | D Trono (EPFL, Lausanne, Switzerland) |
RRID: Addgene_35002 | Lentiviral packaging vector |
| Recombinant DNA reagent |
pVAE2AE4-5 | D Grimm (University of Heidelberg, Germany); Matsushita et al. (1998) |
Adenoviral helper plasmid | |
| Recombinant DNA reagent |
pVpr.IN.eGFP | A Cereseto (CIBIO, Mattareo, Italy); Albanese et al. (2008) |
Plasmid expressing Vpr.IN.eGFP fusion | |
| Software, algorithm |
Autoquant X3 | Media Cybernetics |
RRID:SCR_002465 | |
| Software, algorithm |
GraphPad Prism | GraphPad | RRID:SCR_002798 | |
| Software, algorithm |
Image J | Image J | RRID:SCR_003070 | |
| Software, algorithm |
Imaris 8.1 | BitPlane AG | RRID:SCR_007370 | |
| Software, algorithm |
Imspector | Abberior Instruments |
RRID:SCR_015249 | |
| Software, algorithm |
KNIME | Konstanz Information Miner |
RRID:SCR_006164 | |
| Software, algorithm |
Volocity | Perkin Elmer | RRID:SCR_002668 |