Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) |
HEK293T | ATCC CRL-3216 | RRID:CVCL_0063 | Purchased from ATCC, not verified further |
| Cell line (Homo sapiens) |
hTERT- RPE-1 |
ATCC CRL-4000 | RRID:CVCL_4388 | Purchased from ATCC, not verified further |
| Antibody | Acetylated tubulin | Sigma-Aldrich | Cat# T6793 RRID:AB_477585 |
1:2000 for IF |
| Antibody | HA | Cell Signaling Technology |
Cat# 3724 RRID:AB_1549585 |
1:2000 WB, 1:1000 IF |
| Antibody | IFT88 | Proteintech Group | Cat# 13967–1-AP RRID:AB_2121979 |
1:200 WB, 1:300 IF |
| Antibody | IFT140 | Proteintech Group | Cat# 17460–1-AP RRID:AB_2295648 |
1:200, WB 1:100 IF |
| Antibody | IFT43 | Proteintech Group | Cat# 24338–1-AP RRID:AB_2749824 |
1:50 IF |
| Antibody | IFT20 | Proteintech Group | Cat# 13615–1-AP RRID:AB_2280001 |
1:200 IF |
| Antibody | TMEM67 | Proteintech Group | Cat# 13975–1-AP RRID:AB_10638441 |
1:50 IF |
| Antibody | RPGRIP1L | Proteintech Group | Cat# 55160–1-AP RRID:AB_10860269 |
1:100 IF |
| Antibody | DYNC2H1 | Proteintech Group | Cat# 55473–1-AP AB_2749823 |
1:100 IF |
| Antibody | DYNC2LI1/LIC3 | Proteintech Group | Cat# 15949–1-AP RRID:AB_2093643 |
1:250 WB, 1:100 IF |
| Antibody | TCTEX1 | Santa Cruz Biotechnology |
Cat# sc-28537 RRID:AB_2093671 |
1:200 WB, 1:100 IF |
| Antibody | Arl13B | Proteintech Group | Cat# 17711–1-AP RRID:AB_2060867 |
1:1000 IF |
| Antibody | TCTN1 | Proteintech Group | Cat# 15004–1-AP RRID:AB_10644442 |
1:100 IF |
| Antibody | Smoothened | Abcam | Cat# ab38686 RRID:AB_882615 |
1:100 IF |
| Antibody | Myc | gift from Harry Mellor |
PMID:20233848 | 1:1000 IF |
| Antibody | WDR60 | Sigma-Aldrich | Cat# HPA021316 RRID:AB_2215577 |
1:300 WB in Figure 1—figure supplement 2 and Figure 9—figure supplement 1 |
| Antibody | WDR60 | Novus | Cat# NBP1-90437 RRID:AB_11023602 |
WB in Figure 9 |
| Antibody | WDR34 | Novus | Cat# NBP1-88805 RRID:AB_11006071 |
1:300 WB |
| Antibody | GAPDH | Abcam | Cat# ab9484 RRID:AB_307274 |
1:1000 WB |
| Antibody | p150glued | BD Labs | Cat# 612709 RRID:AB_399948 |
1:1000 WB |
| Antibody | LIS1 | Bethyl | Cat# A300-409A RRID:AB_2159907 |
1:1000 WB |
| Antibody | dic74 | Millipore | Cat# MAB1618 RRID:AB_2246059 |
1:1000 WB |
| Antibody | NUDCD3 | Atlas Antibodies | Cat# HPA019136 RRID:AB_1852370 |
1:350 WB |
| Antibody | Gamma-Tubulin | Sigma-Aldrich | Cat# T5326, RRID:AB_532292 |
|
| Antibody | Anti-HA-Agarose | Sigma-Aldrich | Cat# A2095 RRID:AB_257974 |
|
| Antibody | Alexa- secondary antibodies |
Invitrogen | 1:1000 | |
| Antibody | HRP-secondaries | Jackson ImmunoResearch |
||
| recombinant DNA reagent |
pSpCas9(BB)−2A-GFP | Addgene | Cat# PX458 PMID:24157548 |
|
| recombinant DNA reagent |
pGEM T Easy vector | PROMEGA | Cat# A1360 | |
| recombinant DNA reagent |
L13-Arl13b-GFP (plasmid) |
gift from Tamara Caspary/Addgene |
Cat# 40879 PMID:21976698 |
|
| recombinant DNA reagent |
IFT20-GFP (plasmid) |
gift from Gregory Pazour/Addgene |
Cat# 45608 PMID:16775004 |
|
| recombinant DNA reagent |
pEGFPN3-SSTR3 (plasmid) |
gifts from Kirk Mykytyn/Addgene |
Cat# 35624 PMID:18256283 |
|
| recombinant DNA reagent |
pEGFPN3-5HT6 (plasmid) |
Addgene | Cat# 35623 PMID:18256283 |
|
| recombinant DNA reagent |
EGFP-Rab8 (plasmid) |
gift from Johan Peränen |
||
| recombinant DNA reagent |
pLVX-Puro-mGFP -WDR34 (plasmid) |
PMID:25205765 |
NM_052844.3, NP_443076 |
|
| recombinant DNA reagent |
WDR60 (cDNA) | Life Technologies | Uniprot: Q8WVS4 ENSG00000126870 |
|
| recombinant DNA reagent |
pLVX-Puro-HA- WDR60 (plasmid) |
this paper | ||
| recombinant DNA reagent |
pLVX-Puro-HA- WDR60 T749M (plasmid) |
this paper | ||
| recombinant DNA reagent |
pLVX-Puro-Myc- WDR60 T749M (plasmid) |
this paper | ||
| recombinant DNA reagent |
pLVX-Puro-HA- WDR60 Q631* (plasmid) |
this paper | ||
| chemical compound, drug |
SAG | Selleckchem | Cat# S7779 | |
| chemical compound, drug |
DAPI | Life Technologies | Cat# D1306 | |
| chemical compound, drug |
Amersham ECL | GE Healthcare | Cat# RPN2106 | |
| chemical compound, drug |
DSP | Thermo Fisher Scientific |
Cat# 22585 | |
| chemical compound, drug |
Protease inhibitors | Millipore | Cat# 539137 | |
| chemical compound, drug |
LDS sample buffer | Life Technologies | Cat# NP007 | |
| chemical compound, drug |
Sample reducing agent |
Life Technologies | Cat# NP007 | |
| chemical compound, drug |
Lipofectamine 2000 |
Life Technologies | Cat# 11668027 | |
| chemical compound, drug |
Lenti-XTM HTX Packaging System |
Clontech | Cat# 631275 | |
| Sequence-based reagent |
WDR34 gRNA sequences (Exon2) |
5'-AGCCTTTCTTC GGAGAGTGG-3' |
||
| Sequence-based reagent |
WDR34 gRNA sequences (Exon3) |
5'-CAGGTGTCTTGT CTGTATAC −3' |
||
| Sequence-based reagent |
WDR60 gRNA sequences (Exon3) |
5'-AGGTGCAGGGA TCCCGACCA-3' |
||
| Software and Algorithms |
FIJI/ImageJ | https://fiji.sc/ | PMID:22743772 PMID:22743772 PMID:22743772 |
|
| Software and Algorithms |
Prism 6 | http://www.graphpad.com | ||
| Software and Algorithms |
Proteome Discoverer software v2.1 |
Thermo Fisher Scientific |