KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse Anti-PKC zeta (H1) | SCBT | SC-17781 |
| Rabbit Anti-PKC zeta (C-20) | SCBT | SC-216 |
| Rabbit Anti-PKC zeta p-410 | SCBT | SC-12894-R |
| Rabbit Anti-PKC zeta p560 | Abcam | Ab62372 |
| Mouse Anti-His (AD1.1.10 monoclonal) | SCBT | SC-53073 |
| Rat Anti-Par-6 (polyclonal custom antibody) | Alpha diagnostic | N/A |
| Rabbit Anti- Par-3 S980 phosphospecific | Daniel St. Johnston Lab | |
| Rat Anti-Mira | Abcam | Ab197788 |
| Rabbit Anti-HA (C29F4) | Cell Signaling Tech. | 3724 |
| Mouse Anti-HA | Covance | MMS-101P |
| Guinea Pig Anti-Baz (polyclonal custom antibody) | C.Q.Doe Lab | N/A |
| Chicken Anti-GFP | Abcam | Ab13970 |
| Donkey Anti-Rat Cy3 | Jackson Immuoresearch Lab. | 712-165-153 |
| Donkey Anti-Rabbit 647 | Jackson Immunoresearch Lab. | 711-605-152 |
| Donkey Anti-Mouse 647 | Jackson Immunoresearch Lab. | 715-605-151 |
| Donkey Anti-Mouse 488 | Jackson Immunoresearch Lab. | 715-545-151 |
| Donkey Anti-Chicken 488 | Jackson Immunoresearch Lab. | 703-545-155 |
| Donkey Anti-Guinea Pig 405 | Jackson Immunoresearch Lab. | 706-475-148 |
| IRDye 680RD Donkey anti-mouse secondary | Licor | 926–68072 |
| IRDye 800CW Goat anti-rat secondary | Licor | 926–32219 |
| Bacterial and Virus Strains | ||
| BL21-DE3 | ||
| TG1 | ||
| Biological Samples | ||
| Chemicals, Peptides, and Recombinant Proteins | ||
| All Par-3 constructs prepared from Drosophila melanogaster transcript variant A: NM_001347807.1 | ||
| MBP – Par-3 (1 – 1464) | This Paper | MBP – Par-3 - WT |
| MBP – Par-3 -His (309 – 991) | This Paper | MBP – Par-3 – PDZ1 – APM – His |
| MBP – Par-3 (393 – 1464) | This Paper | MBP – Par-3 – ΔOD -PDZ1 |
| MBP – Par-3 (1 – 1464 Δ 437–533) | This Paper | MBP – Par-3 – ΔPDZ2 |
| MBP – Par-3 (1 – 1464 Δ 616 – 741) | This Paper | MBP – Par-3 – ΔPDZ3 |
| MBP – Par-3 (1 – 1464 Δ 969 – 987) | This Paper | MBP – Par-3 – ΔAPM |
| MBP – Par-3 (309 – 1464) | This Paper | MBP – Par-3 – PDZ1 – COOH |
| MBP – Par-3 (1– 987) | This Paper | MBP – Par-3 – NH2 term – APM |
| MBP – Par-3 (1–1464) (S980A) | This Paper |
MBP-Par-3
S980A |
| His – Dr - Par-3 (437 – 741) | This Paper | His – DrPar-3 – PDZ2 |
| His – Human-Par-3 (454 – 549) (Q9BYG5) | This Paper | His – HuPar-3 – PDZ2 |
| His – Hydra - Par-3 (370 – 463) (T2M6J3) | This Paper | His – HyPar-3 – PDZ2 |
| His – Trichoplax - Par-3 (250–340) (A0A369RWF0) | This Paper | His – TrPar-3 – PDZ2 |
| GST-Dr – aPKC (601 – 606) (A1Z9X0) | This Paper | GST-DraPKC – PBM |
| GST-Dr – aPKC (601 – 606 -V606A) (A1Z9X0) | This Paper | GST-DraPKC – PBM (V606A) |
| GST-Human – aPKC (591–596) (P41743) | This Paper | GST-HuaPKC – PBM |
| GST-Hydra – aPKC (574–579) (T2MGA0) | This Paper | GST-HyaPKC – PBM |
| GST-Trichoplax – aPKC (404 – 410)(A0A369S098) | This Paper | GST-TraPKC – PBM |
| aPKC (1 – 606) | This Paper | aPKC |
| aPKC (1 – 606) (V606A) | This Paper | aPKC (V606A) |
| aPKC ΔPBM (1 – 600) | This Paper | aPKC ΔPBM |
| His – Par-6 (1 – 351) | This Paper | His – Par-6 |
| His – Par-6 ΔPBM (1 – 343) | This Paper | His – Par-6 ΔPBM |
| 293fectin | ThermoFisher | 12347019 |
| Freestyle 293 expression Medium | ThermoFisher | 12338026 |
| Opti-Mem | ThermoFisher | 3198588 |
| HisPur cobalt resin | ThermoFisher | 89965 |
| Amylose Resin | NEB | E8021L |
| Glutathione Resin | GoldBio | G250–100 |
| Source Q anion exchange resin | GE Healthcare | 17-1275-01 |
| Bolt 12% Bis-Tris Gels | ThermoFisher | NW00125BOX |
| TMB Substrate | ThermoFisher | N301 |
| Odyssey blocking buffer (TBS) | LiCor | 927–50000 |
| Schneider’s Insect Medium (SIM) | Sigma | S0146 |
| SlowFade Diamond Antifade Mountant with DAPI | Invitrogen | S36964 |
| Critical Commercial Assays | ||
| Deposited Data | ||
| Experimental Models: Cell Lines | ||
| Human Embryonic Kindey cells (HEK293) | ThermoFisher | R79007 |
| Experimental Models: Organisms/Strains | ||
| D. melanogaster: ;Worniu-Gal4 | Bloomington Drosophila Stock Center | RRID:BDSC_56553 |
| D. melanogaster: ;;3xHA-aPKC 1–606 | This Paper | N/A |
| D. melanogaster: ;;3xHA-aPKC V606A | This Paper | N/A |
| D. melanogaster: ; FRT-G13, aPKCK06403/CyO | C.Q. Doe Lab | N/A |
| D. melanogaster: elav-Gal4, UAS-mCD8:GFP, hs:flp; FRT-G13, tubPGal80 | Bloomington Drosophila Stock Center | RRID:BDSC_5145 |
| Oligonucleotides | ||
| Recombinant DNA | ||
| pCMV mammalian expression plasmid | ThermoFisher | 10586014 |
| pMal C4X bacterial expression plasmid | Addgene | 75288 |
| pGex 4Ti bacterial expression plasmid | Amersham | 27458001 |
| pUASTattB fly cloning and transformation plasmid | Addgene | EF362409.1 |
| Software and Algorithms | ||
| ImageJ | ||
| GraphPad | ||
| Other | ||
| PD10 Desalting columns | 95017–001 | |
| VivaSpin 20 sample concentrators MWCO 30kD | VWR | 95056–130 |
| VivaSpin 20 sample concentrators MWCO 10kD | VWR | 95056–128 |
| Shaker Flasks – 125mL | VWR | 89095–258 |