REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Mouse monoclonal anti-NSD1 | Antibodies, Inc. | Cat#75-280; RRID: AB_11001827 |
Mouse monoclonal anti-NSD2 29D1 | Abcam | Cat#ab75359; RRID: AB_1310816 |
Rabbit monoclonal anti-NSD3 D4N9N | Cell Signaling | Cat# 92056, RRID:AB_2800178 |
Rabbit monoclonal anti-BRG1 | Cell Signaling | Cat#49360; RRID: AB_2728743 |
Rabbit monoclonal anti-BAF155 | Cell Signaling | Cat# 11956; RRID: AB_2797776 |
Rabbit monoclonal anti-SMARCB1 | Cell Signaling | Cat#91735; RRID: AB_2800172 |
Mouse monoclonal anti-EZH2 | Active Motif | Cat# 39875; RRID: AB_2561022 |
Rabbit monoclonal anti-EZH2 | Cell Signaling | Cat#5246; RRID: AB_10694683 |
Rabbit monoclonal anti-SUZ12 | Cell Signaling | Cat#3737; RRID: AB_2196850 |
Mouse monoclonal anti-EED | Millipore Sigma | Cat#05-1320; RRID: AB_1586999 |
Rabbit polyclonal anti-EED | Millipore Sigma | Cat#09-774; RRID: AB_1587000 |
Rabbit monoclonal anti-BMI1 | Cell Signaling | Cat#6964; RRID: AB_10828713 |
Rabbit monoclonal anti-RING1B | Cell Signaling | Cat#5694; RRID: AB_10705604 |
Mouse monoclonal anti-b-Actin | Sigma Aldrich | Cat#A5441; RRID: AB_476744 |
Rabbit polyclonal anti-HSP90 | Cell Signaling | Cat# 4874, RRID:AB_2121214 |
Rabbit monoclonal anti-KDM2A | Abcam | Cat#ab191387 |
Mouse monoclonal anti-Lamin A/C | Cell Signaling | Cat# 4777, RRID:AB_10545756 |
Rabbit monoclonal anti-Histone H3 | Cell Signaling | Cat#4620; RRID: AB_1904005 |
Rabbit polyclonal H3K27ac | Abcam | Cat#ab4729; RRID: AB_2118291 |
Rabbit monoclonal H3K27me3 C36B11 | Cell Signaling | Cat#9733; RRID: AB_2616029 |
Rabbit polyclonal H3K36me2 | Abcam | Cat#ab9049; RRID: AB_1280939 |
Rabbit monoclonal Rabbit Control IgG | Cell Signaling | Cat#3900; RRID: AB_1550038 |
Normal mouse IgG | Santa Cruz | Cat#sc-2025; RRID: AB_737182 |
Drosophila spiked in antibody | Active Motif | Cat# 61686; RRID: AB_27370 |
Monoclonal mouse anti-Rabbit IgG, light chain specific | Jackson | Cat#211-032-171; RRID: AB_2339149 |
Goat anti-mouse IgG, light chain specific | Jackson | Cat#115-035-174; RRID: AB_2338512 |
Chemicals, peptides, and recombinant proteins | ||
GSK126 | ChemieTek | Cat#CT-GSK126 |
TC-E5002 | Tocris | Cat#5089 |
Vincristine (Tocris #1257 | Tocris | Cat#1257 |
Doxorubicin (CarboSynth #AC15377 | CarboSynth | Cat#AC15377 |
D-actinomycin (ApexBio #A4448 | ApexBio | Cat#A4448 |
Etoposide (AG Chemicals #E2004 | AG Chemicals | Cat#E2004 |
Dimethylsulfoxide (DMSO) | ATCC | Cat#4-X |
Critical commercial assays | ||
DirectZol Mini Prep Plus kit | Zymo Research | Cat#R2070 |
Quant-iT RiboGreen assay | ThermoFisher | Cat#R11490 |
Epicypher Cutana Cut&Run Kit v2.0 | Epicypher | Cat# 14-1048 |
Covaris True- ChIP kit | Covaris | Cat#520154 |
Drosophila chromatin spike in antibody | Active Motif | Cat#61686 |
Dynabeads™ Protein G for Immunoprecipitation | ThermoFisher | Cat#10004D |
RNase A | ThermoFisher | Cat#EN0531 |
Ptoteinase K | ThermoFisher | Cat#4333793 |
Nuclease | ThermoFisher | Cat#88702 |
Cell titer aqueous reagent | Promega | Cat#G3582 |
CellTiter-Glo | Promega | Cat#G9241 |
Halt™ Protease and Phosphatase Inhibitor Cocktail (100X) | ThermoFisher | Cat# 78446 |
Deposited data | ||
RNAseq data | This study | GSE90631 |
ChIP-seq data | This study | GSE90631 |
Cut&Run data | This study | GSE90631 |
Uncropped immunoblot data | This study | Mendeley data doi: 10.17632/fvzm88tm3v.2 |
Experimental models: Cell lines | ||
G401 | ATCC | CVCL_0270 |
G402 | ATCC | CVCL_1221 |
A549 | ATCC | CVCL_0023 |
TOV21G | ATCC | CVCL_3613 |
HFF-1 | ATCC | CVCL_3285 |
HEK293T | ATCC | CVCL_0063 |
Pfeiffer | ATCC | CVCL_3326 |
BT16 | Laboratory of Dr. C. David James | CVCL_M156 |
TTC709 | Laboratory of Dr. B.E. Weissman. | CVCL_8007 |
Oligonucleotides | ||
sgRNA for NSD1 knockout: SP185.g12 5- UCUCCAACUUCAUACUUCAG-3 |
This study | NA |
Forward primer for Next Generation Sequence (NGS) of the NSD1 gene and assessment of indels: 5-CATACATTGCTTTTTCAGAAGGCT-3 |
This study | NA |
Reverse primer for Next Generation Sequence (NGS) of the NSD1 gene and assessment of indels: 5- ACACCAATTCATTCACAAAATGTTC-3 |
This study | NA |
sgRNA for NSD2 knockout: g7 5-CCAAAGUGUCGGGUUACCCU-3 |
This study | NA |
Forward primer for Next Generation Sequence (NGS) of the NSD2 gene and assessment of indels: 5- AGCCTGGTGAATCCTTGGTCTTCTGG-3 |
This study | NA |
Reverse primer for Next Generation Sequence (NGS) of the NSD2 gene and assessment of indels: 5-GCTGCAAAAATGAACCATAGCAGGGC-3 |
This study | NA |
sgRNA for NSD3 knockout: g16 5-AAACUGGACAUGAUAUUCUC-3 |
This study | NA |
Forward primer for Next Generation Sequence (NGS) of the NSD3 gene and assessment of indels: 5-ACAGGTGCCCGAGAATATCA-3 |
This study | NA |
Reverse primer for Next Generation Sequence (NGS) of the NSD3 gene and assessment of indels: 5- ACGATTAAAACAGCTGAAAGGGG-3 |
This study | NA |
Recombinant DNA | ||
Plasmid: pLKO_005-shNSD1-1 | Sigma | TRCN0000238371 |
Plasmid: pLKO_005-shNSD1-2 | Sigma | TRCN0000238372 |
Plasmid: pLKO.1-shKDM2A-1 | Sigma | TRCN0000022000 |
Plasmid: pLKO_005-shKDM2A-2 | Sigma | TRCN0000359598 |
Plasmid: pLV[Exp]-Bsd-CMV>Tet3G | VectorBuilder | 180621-1181ctz |
Plasmid: pLV[Exp]-Puro-TRE3G>EGFP | VectorBuilder | 180621-1204vsc |
Plasmid: pLV[Exp]-Puro-TRE3G>V5/hSMARCB1 | VectorBuilder | 180622-1078xvz |
Plasmid: pVSV-G | Addgene | Addgene #138479 |
Plasmid: psPAX2 | Addgene | Addgene #12260 |
Software and algorithms | ||
Prism 7.0d | GraphPad | www.graphpad.com |
Image J 2.1.0/1.53c | NIH | https://imagej.nih.gov/ij |
Integrative Genomics Viewer (IGV_2.6.2) | The Broad Institute | https://software.broadinstitute.org/software/igv/ |