Table 1 List of materials used in this study
Material | Company | CAT# |
|---|---|---|
Antibodies | ||
Mouse monoclonal anti-Actin | Thermo Fisher Scientific (Invitrogen) | MA1-744 |
Rabbit anti-Gag | D. Garfinkel | N/A |
Mouse monoclonal anti-Ty1-Tag | Sigma-Aldrich | SAB4800032 |
Rabbit peroxidase anti-peroxidase | Sigma-Aldrich | P1291 |
Mouse monoclonal anti-PABP1 | EnCor Biotech | MCA-1G1 |
Rabbit anti-Trf4 | D. Tollervey | N/A |
Mouse monoclonal anti-RNA Pol II (Phosphorylated Serine 5) | AbCam | ab5408 |
Mouse monoclonal S9.6 (anti-RNA–DNA Hybrid) | KeraFast | ENH002 |
Mouse monoclonal anti-MYC | Covance research | MMS-150R-500 |
Mouse monoclonal anti-MYC HRP | Thermo Fisher Scientific (Invitrogen) | R951-25 |
Rabbit polyclonal anti-acetyl histone H3 | Millipore Sigma | 06-599 |
Rabbit polyclonal anti-histone H3 | AbCam | Ab1791 |
Rabbit polyclonal Anti-ATXN2 | Proteintech | 21776-1-AP |
Mouse monoclonal anti-ɑ-tubulin | Sigma | T6074 |
Chemicals | ||
|---|---|---|
1,10-phenanthroline | Sigma-Aldrich | 131377-25G |
DNase I (Bovine Pancreas) | Sigma-Aldrich | D7291 |
SensiFAST SYBR No-ROX Kit | BIOLINE | BIO-98050 |
Pierce ECL western blotting substrate | Thermo Fisher Scientific | 32106 |
cOmplete protease inhibitor (Roche) | Sigma-Aldrich | 11697498001 |
IgG sepharose 6 fast flow beads | Sigma-Aldrich | GE17-0969-01 |
Protein-A sepharose beads | GE Healthcare | 17078001 |
Lipofectamine RNAiMAX | Thermo Fisher Scientific | 13778150 |
LipoD293 | Signagen | SL100668 |
SeaKem LE agarose | Lonza | 50001 |
Commercial Assays | ||
|---|---|---|
Amersham megaprime DNA labeling system | GE Healthcare | RPN1606 |
M-MLV reverse transcriptase kit | Thermo Fisher Scientific (Invitrogen) | 28025013 |
RNeasy mini kit | Qiagen | 74104 |
KAPA2G Fast HotStart PCR Kit | KAPA Biosystems | KK5517 |
Oligonucleotides | ||
|---|---|---|
siRNA targeting ATXN2 | Ambion | 4390843 |
siRNA control | Ambion | 4392420 |
TYA: ATCTATGATTCCGTATACAC and AGGATGAATCAGTAAATGTA | This study | N/A |
TYB: AGAATACCGAGGAATCTATCATCGC and AGTCACCAATACCACCCAAACTG | This study | N/A |
TYB-2: GACCGTGGTTCTGAGTATACT and TTCGGTAAACCACTACATTG | This study | N/A |
Ty1His3: GGCCGTGCGTGGAGTAAAAA and AAGAAAATGCGGGATCATCTC | This study | N/A |
Integration US tRNAGLY: GTGATGACAAAACCTCTTCCG and GGCAACGTTGGATTTTACCAC | N/A | |
Integration US tRNATYR: GTGATGACAAAACCTCTTCCG and AGTCTTGCGCCTTAAACCAA | 65 and this study | N/A |
TEL1: CGGATTTCTGACGATATGGAC and ACCAACGTACTGAAGGTATCC | N/A | |
Cup1: TGAAGGTCATGAGTGCCAAT and TTCGTTTCATTTCCCAGAGCA | N/A | |
Act1: GCCTTCTACGTTTCCATCCA and GGCCAAATCGATTCTCAAAA | N/A | |
Pbp1: AAACCGATCAGCAAAACCCC and CCGGTAGATGATTGGCCTTG | This paper | N/A |
IGS P1: CCGGGGCCTAGTTTAGAGAG and ACCCATCTTTGCAACGAAAA | N/A | |
IGS P2: AGGGCTTTCACAAAGCTTCC and TCCCCACTGTTCACTGTTCA | N/A | |
IGS P3: TGATGATGGCAAGTTCCAGA and CTTATTCCTTCCCGCTTTCC | N/A | |
IGS P4: GGAAAGCGGGAAGGAATAAG and CGATTCAGAAAAATTCGCACT | N/A | |
IGS P5: GTTGGTTTTGGTTTCGGTTG and TCGCCGAGAAAAACTTCAAT | N/A | |
25S rDNA: GGGAATGCAGCTCTAAGTGG and ATGGATTTATCCTGCCACC | This study | N/A |
ATXN2: CTCCTCGGTGGTCGCGGCGACCTC and CTCTTTTTGCATAACTGGAGTCC | N/A | |
GAPDH: TGAAGGTCGGAGTCAACGGATTTGG and GGAGGCCATGTGGGCCATGAG | N/A | |
RPLP0: GGCGACCTGGAAGTCCAACT and CCATCAGCACCACAGCCTTC | N/A | |
LINE-1: TGCGGAGAAATAGGAACACTTTT and TGAGGAATCGCCACACTGACT | N/A | |
HERV-H: CACGTTTTATCCGTGGACCC and AGGCATCCCTGCAATGATTAA | N/A | |
Alu: TAGCCAGGTGTGGTGACTTG and AATGGTACGATCTFGGCTCA | N/A | |
Recombinant DNA | ||
|---|---|---|
p426 Empty GPD URA Vector (pKM55) | ATCC | ATCC87361 |
p426 GPD Hsp104 URA (pKM302) | Duennwald lab | |
p416 GAL Pbp1-1Q URA (pKM309) | This study | N/A |
p416 GAL Pbp1-5Q URA (pKM310) | This study | N/A |
p416 GAL Pbp1-15Q URA (pKM311) | This study | N/A |
p416 GAL Pbp1-30Q URA (pKM312) | This study | N/A |
p416 GAL Pbp1-40Q URA (pKM313) | This study | N/A |
p416 Empty GAL URA Vector (pKM279) | This study | N/A |
ATNX2-23Q-Halo (pKM286) | N/A | |
ATXN2-31Q-Halo (pKM288) | N/A | |
ATXN2-39Q-Halo (pKM291) | N/A | |
pRS303 Empty Vector HIS | ATCC | ATCC77138 |
pRS303 Pbp1-1Q-TAP HIS | This study | N/A |
pRS303 Pbp1-5Q-TAP HIS | This study | Thermo Fisher Scientific |
pRS303 Pbp1-15Q-TAP HIS | This study | Thermo Fisher Scientific |
pRS303 Pbp1-30Q-TAP HIS | This study | Thermo Fisher Scientific |
pRS303 Pbp1-40Q-TAP HIS | This study | Thermo Fisher Scientific |
pLTR:Gag1-401-GFP LEU (pKM353) | Curcio lab | |
pRS316 Nop1-CFP URA (pKM354) | This study | N/A |
