REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Rabbit Anti-Human SAMHD1 Polyclonal Antibody | Proteintech | Cat# 12586-1-AP, RRID:AB_2183496 |
Mouse Anti-Actin, beta Monoclonal Antibody | Abcam | Cat# ab6276, RRID:AB_2223210 |
Rabbit polyclonal CDK1 Antibody | Bethyl | Cat# A303-664A, RRID:AB_11204758 |
Mouse Anti-BM28 Monoclonal Antibody | BD Biosciences | Cat# 610701, RRID:AB_398024 |
Phospho-SAMHD1 (Thr592) (D7O2M) Rabbit mAb antibody | Cell Signaling Technology | Cat# 89930, RRID:AB_2800147 |
Mouse Anti-P21 Monoclonal antibody, Unconjugated, Clone f-5 | Santa Cruz Biotechnology | Cat# sc-6246, RRID:AB_628073 |
IRF-3 (D6I4C) XP® antibody | Cell Signaling Technology | Cat# 11904, RRID:AB_2722521 |
NFkappaB p65 (F-6) antibody | Santa Cruz Biotechnology | Cat# sc-8008, RRID:AB_628017 |
Purified anti-H2A.X Phospho (Ser139) antibody | BioLegend | Cat# 613402, RRID:AB_315795 |
53BP1 antibody | BD Biosciences | Cat# 612522, RRID:AB_2206766 |
Anti-IFNα/β Receptor | PBL Interferon Source | N/A |
Mouse IgG2A Isotype Control (Clone 20102) antibody | R and D Systems | Cat# MAB003, RRID:AB_357345 |
Rat Anti-IL-6 Monoclonal Antibody, Unconjugated, Clone MQ2-13A5 | BD Biosciences | Cat# 554543, RRID:AB_398568 |
Mouse Anti-IL-8 Monoclonal Antibody, Unconjugated, Clone G265-5 | BD Biosciences | Cat# 554716, RRID:AB_395526 |
Mouse Anti-TNF Monoclonal Antibody, Unconjugated, Clone MAb1 | BD Biosciences | Cat# 551220, RRID:AB_394098 |
Rabbit Anti-Human TICAM1 Polyclonal Antibody, Unconjugated | GeneTex | Cat# GTX104744, RRID:AB_1241389 |
TBK1 Antibody | Bethyl | Cat# A300-093A, RRID:AB_2303002 |
Bacterial and Virus Strains | ||
VSV-G HIV-1 GFP virus | Besnier et al., 2002 | N/A |
SIVmac Virus like particles | Goujon et al., 2008, Reinhard et al., 2014 | N/A |
Biological Samples | ||
PBMC from HIV seronegative donors | This study | N/A |
Chemicals, Peptides, and Recombinant Proteins | ||
AB Human Serum | Sigma | #H4522 |
FBS | Biosera | N/A |
BX795 | APEXBIO | #A8222 |
Ruxolitinib | Cell guidance system | #SM87 |
Recombinant human IFN-b | Peprotech | #300-02BC |
Recombinant human Tenascin C | R and D | #3358-TC-050 |
Polymyxin B | Sigma | #P4932 |
LPS | InvivoGen | #tlrl-prslps |
TAK242 | Milllipore | #614316 |
Recombinant Flagellin protein | Abcam | #Ab201366 |
DMEM, high glucose, pyruvate | Invitrogen | #41966052 |
RPMI 1640 medium | Invitrogen | #21875091 |
Opti-MEM | Invitrogen | #31985047 |
Fugene HD transfection reagent | Promega | #E2311 |
DAPI | Sigma | #10236276001 |
PhosSTOP | Sigma | #4906845001 |
Fast SYRB green master mix | ThermoFisher | #4385610 |
Total RNA Purification Kit | Norgen | #17200 |
Critical Commercial Assays | ||
CellRox | Molecular Probes | #C10444 |
NuPAGE 4-12% Bis-Tris Protein Gel | Invitrogen | #NP0322BOX |
pHrodo E.coli BioParticles | ThermoFisher | #P35361 |
SuperScript III First-Strand Synthesis System | ThermoFisher | #18080051 |
Amersham ECL Prime Western Blotting Detection Reagent | GE Healthcare | #RPN2232 |
Experimental Models: Cell Lines | ||
293T | Laboratory of G. Towers (UCL) | N/A |
Oligonucleotides | ||
CDK1 FWD 5′TGAGGAACGGGGTCCTCTAA 3′ |
Invitrogen | N/A |
CDK1 R 5′ A TGGCT ACCACTTGA CCTGT 3′ |
Invitrogen | N/A |
CDK2 F 5′ AAGTTGACGGGAGAG GTGGT 3′ |
Invitrogen | N/A |
CDK2 R 5′ TGATGAGGGGAAGAG GAATG 3′ |
Invitrogen | N/A |
CDK4 F 5′ CAGATGGCACTTACACCCGT 3′ | Invitrogen | N/A |
CDK4 R 5′ CAGCCCAATCAGGTC AAAGA 3′ |
Invitrogen | N/A |
CDK6 F 5′ CGTGGTCAGGTTGTT TGATGT 3′ |
Invitrogen | N/A |
CDK6 R 5′ CGGTGTGAATGAAGAA AGTCC 3′ |
Invitrogen | N/A |
Cyclin A2 F 5′ AAGACGAGACGGGTTGC 3′ | Invitrogen | N/A |
Cyclin A2 R 5′ GGCTGTTTACTGTTTGCTTTCC 3′ |
Invitrogen | N/A |
Cyclin B1 F 5′ TTCTGGATAATGGTGAATGGAC 3′ |
Invitrogen | N/A |
Cyclin B1 R 5′ ATGTGGCATACTTGTTCTTGAC 3′ |
Invitrogen | N/A |
Cyclin D1 F 5′ AGATGAAGGAGACCATCCCCC 3′ |
Invitrogen | N/A |
Cyclin D1 R 5′ CCACTTGAGCTTGTTCACCA 3′ |
Invitrogen | N/A |
Cyclin D3 F 5′ GGCCGGGGACCGAAACT 3′ | Invitrogen | N/A |
Cyclin D3 R 5′CAGTGGCGAAGTGTTTACAAAGT 3′ |
Invitrogen | N/A |
Cyclin E1 F 5′ CCGGTATATGGCGACACAAG 3′ |
Invitrogen | N/A |
Cyclin E1 R 5′ TACGCAAACTGGTGCAACTT 3′ |
Invitrogen | N/A |
Cyclin E2 F 5′ TCTCCTGGCTAAATCTCTTTCTCC 3′ |
Invitrogen | N/A |
Cyclin E2 R 5′ ACTGTCCCACTCCAAACCTG 3′ |
Invitrogen | N/A |
E2F1 F 5′ TGCCAAGAAGTCCAAGA ACCA 3′ |
Invitrogen | N/A |
E2F1 R 5′ GTCAACCCCTCAAGCCGTC 3′ | Invitrogen | N/A |
E2F4 F 5′ CGGACCCAACCCTTCT ACCT 3′ | Invitrogen | N/A |
E2F4 R 5′ GGGGCAAACACTTCTGAGGA 3′ | Invitrogen | N/A |
E2F7 F 5′ CCTTTAGCCCACCCAGTATTT 3′ | Invitrogen | N/A |
E2F7 R 5′ A TCCCTCTCTGACCCTGACC 3′ | Invitrogen | N/A |
MCM2 F 5′CACCCGAAGCTCAACCA GAT 3′ |
Invitrogen | N/A |
MCM2 R 5′ATCATGGACTCGATGTG CCG 3′ |
Invitrogen | N/A |
RB1 F 5′AAAGGACCGAGAAGGACCA 3′ | Invitrogen | N/A |
RB1 R 5′AAGGCTGAGGTTGCTTGTGT 3′ | Invitrogen | N/A |
WIF1 F 5′TCTGTTCAAAGCCTGTCTGC 3′ | Invitrogen | N/A |
WIF1 R 5′ACATTGGCATTTGTTGGGTT 3′ | Invitrogen | N/A |
p14 F 5′GAGTGAGGGTTTTCGTGGTTC 3′ | Invitrogen | N/A |
p14 R 5′ACGGGTCGGGTGAGAGTG 3′ | Invitrogen | N/A |
p16 F 5′ CGGCTGACTGGCTGGC 3′ | Invitrogen | N/A |
p16 R 5′ GGGTCGGGTGAGAGTGG3′ | Invitrogen | N/A |
p21 F 5′GCCGAAGTCAGTTCCTTGTG 3′ | Invitrogen | N/A |
p21 R 5′TCGAAGTTCCATCGCTCACG 3′ | Invitrogen | N/A |
p27 F 5′A TGTTTCAGACGGTTCCCCA 3′ | Invitrogen | N/A |
p27 R 5′TCCAACGCTTTTAGAGGCAG 3′ | Invitrogen | N/A |
p53 F 5′AAGTCTAGAGCCACCGTCCA 3′ | Invitrogen | N/A |
p53 R 5′TTTCAGGAAGTAGTTTCCATAGGT 3′ |
Invitrogen | N/A |
CXCL10 F 5′ TGGCATTCAAGGAGTACC TC 3′ |
Sigma | N/A |
CXCL10 R 5′ TTGTAGCAATGATCTCAACACG 3′ |
Sigma | N/A |
ISG56 F 5′CCT CCT TGG GTT CGT CTA CA 3′ |
Sigma | N/A |
ISG56 R 5′GGC TGA TAT CTG GGT GCC TA 3′ |
Sigma | N/A |
ISG54 F 5′CAGCTGAGAATTGCACTGC AA 3′ |
Sigma | N/A |
ISG54 R 5′CGT AGGCTGCTCTCCAAG GA 3′ |
Sigma | N/A |
MxA F 5′ATC CTG GGA TTT TGG GGC TT 3′ |
Sigma | N/A |
MxA R 5′CCG CTT GTC GCT GGT GTC G 3′ | Sigma | N/A |
SAMHD1 F 5′TTGTGCTAGAGATAAGGAAGTTGG 3′ |
Invitrogen | N/A |
SAMHD1 R 5′TGTGTTGATAAGCTCTACGGTG 3′ |
Invitrogen | N/A |
GAPDH F 5′ACC CAG AAG ACT GTG GAT GG 3′ |
Sigma | N/A |
GAPDH R 5′TTC TAG ACG GCA GGT CAG GT 3′ |
Sigma | N/A |
ON-TARGETplus Human SAMHD1 siRNA | Dharmacon | #L-013950-01 |
Control siRNA | Santa Cruz | #sc-37007 |
TBK1 siRNA | Santa Cruz | #sc-39058 |
TICAM1 siRNA | OriGene | #SR315629 |
Recombinant DNA | ||
GFP-encoding genome CSGW | Laboratory of G. Towers | N/A |
p8.91 | Laboratory of G. Towers | N/A |
pMDG | Laboratory of G. Towers | N/A |
SIVmac packaging plasmid encoding accessory genes |
Laboratory of G. Towers | N/A |
pCDNA Vpx E16A | Laboratory of J. Luban, Reinhard et al., 2014 | N/A |
Software and Algorithms | ||
ImageJ | https://imagej.nih.gov/ij/ | |
Other | ||
Amersham Hybond P 0.45 PVDF blotting membrane |
GE Healthcare | #10600023 |