KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| Mouse monoclonal anti-α-tubulin antibody (DM1A) | Sigma Aldrich | Cat#T6199; RRID: AB_477583 |
| Rat monoclonal anti-α-tubulin antibody (YOL1/34) | Abcam | Cat#ab6161; RRID: AB_305329 |
| Mouse monoclonal anti-acetylated α-tubulin antibody (6-11B-1) | Sigma Aldrich | Cat#T7451; RRID: AB_609894 |
| Rat monoclonal anti-α-tubulin antibody (YL1/2) | Abcam | Cat#ab6160; RRID: AB_305328 |
| Mouse monoclonal anti-polyglutamylation antibody | AdipoGen | Cat#AG-20B-0020; RRID: AB_2490210 |
| AF555 goat anti-mouse highly cross-adsorbed secondary antibody | Thermo Fisher | Cat#A21424; RRID: AB_141780 |
| AF488 Goat anti-mouse highly cross-adsorbed secondary antibody | Thermo Fisher | Cat#A32723; RRID: AB_2633275 |
| AF647 Goat anti-mouse highly cross-adsorbed secondary antibody | Thermo Fisher | Cat#A21236; RRID: AB_2535805 |
| AF647 Goat anti-rat highly cross-adsorbed secondary antibody | Thermo Fisher | Cat#A48265; RRID: AB_2895299 |
|
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| Bacterial and virus strains | ||
|
| ||
| Aerobacter aerogenes | Chandler Fulton | N/A |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Difco Antibiotic Medium 3 | Fisher | Cat#DF0243-17-8 |
| Gibco Bacto Peptone | Fisher | Cat#DF0118-17-0 |
| Gibco Yeast Extract | Fisher | Cat#B11929 |
| TRIZol Reagent | Thermo Fisher | Cat#15596026 |
| Benomyl | Sigma Aldrich | Cat#45339 |
| Colchicine | Sigma Aldrich | Cat#C9754 |
| Nocodazole | Sigma Aldrich | Cat#M1404 |
| Oryzalin | Sigma Aldrich | Cat#36182 |
| Paclitaxel | Sigma Aldrich | Cat#T7402 |
| Plinabulin | Sigma Aldrich | Cat#ADV947322154 |
| Vinblastine | Sigma Aldrich | Cat#V1377 |
| DMSO (anhydrous) | Sigma Aldrich | Cat#D2650 |
| NP-40 Alternative | Sigma Aldrich | Cat#492016 |
| Detector Block | SeraCare | Cat#5920-0004 |
| Triton-X 100 | Promega | Cat#H5142 |
| cold water fish skin gelatin | Sigma Aldrich | Cat#G7765 |
| Tubulin Tracker Deep Red | Thermo Fisher | Cat#T34077 |
| MitoTracker Red CMXRos | Thermo Fisher | Cat#M7512 |
| AF488 Phalloidin | Thermo Fisher | Cat#A12379 |
| Prolong Gold mounting medium with DAPI | Thermo Fisher | Cat#P36935 |
|
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| Critical commercial assays | ||
|
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| Power SYBR Green qPCR Master Mix | Thermo Fisher | Cat#4368706 |
| SuperScript IV First-Strand Synthesis kit | Thermo Fisher | Cat#18091200 |
|
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| Experimental models: Organisms/strains | ||
|
| ||
| Naegleria gruberi: NEG: wildtype | Chandler Fulton | ATCC 30223 |
| Naegleria gruberi: NEG-M: wildtype | Chandler Fulton | ATCC 30224 |
|
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| Oligonucleotides | ||
|
| ||
|
N. gruberi GAPDH (JGI ID: 53883) qPCR primers: Forward 5’-TGGC TCCAATTGCTGCTGTTT-3’ and reverse 5’-CCTTAGCAGCACCAG TTGAAGA-3’ |
This work | N/A |
|
N. gruberi G Protein (JGI ID: 77952) qPCR primers: Forward 5’-ACGGTTGGGTCACTTGTTTGTCC-3’ and reverse 5’-GAGCGTGACCAGTGAGGGATC-3’ |
This work | N/A |
|
N. gruberi mitotic α-tubulin (JGI ID: 58607) qPCR primers: Forward 5’-GGTCCTTGATGTGTGCCGAAC-3’ and reverse 5’-TTAGCAGCATCTTCACGACCAGT-3’ |
This work | N/A |
|
N. gruberi mitotic α-tubulin (JGI ID: 55745) qPCR primers: Forward 5’-CACACACAAAATGAGAGAAGTCGTC-3’ and reverse 5’-TTCCATGTTCAGCACAGAATAATTC-3’ |
This work | N/A |
|
N. gruberi mitotic β-tubulin (JGI ID: 55748) qPCR primers: Forward 5’-AACCAACACTGCTTCTCCACTCG-3’ and reverse 5’-TCTGGACGGAATAATTGACCTTGG-3’ |
This work | N/A |
|
N. gruberi mitotic β-tubulin (JGI ID: 55900) qPCR primers: Forward 5’-GGTTGCTGGTGTCATGTCTGGTG-3’ and reverse 5’-GCAGCCAAAGGAGCAGAACCAA-3’ |
This work | N/A |
|
| ||
| Software and algorithms | ||
|
| ||
| PhyloTOL | 68 | N/A |
| PASTA multiple sequence aligner | 69 | N/A |
| IQ-Tree 2 v.1.16.2 | 70 | RRID: SCR_017254 |
| ITOL v4 | 71 | RRID: SCR_018174 |
| ClustalOmega | 72 | RRID: SCR_001591 |
| AL2CO | 73 | N/A |
| PyMOL v2.4.1 | Schrödinger, LLC | RRID: SCR_000305 |
| NIS Elements with Advanced Research Package | Nikon Instruments | RRID: SCR_014329 |
| Autoquant X3 vX3.1.3 | Media Cybernetics | RRID: SCR_002465 |
| Fiji | 74 | RRID: SCR_002285 |
| ChimeraX | 75 | RRID: SCR_015872 |
| GraphPad Prism v8 | GraphPad | RRID: SCR_002798 |
| Custom scripts to analyze spindle twist | 76 | N/A |
| StepOne v2.3 | Thermo Fisher | RRID: SCR_014281 |