KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse anti-SEPT9 | EMD Millipore | Cat#MABE992 |
| Rabbit anti-septin 9 | PTG labs | Cat#10769 – IAP; RRID:AB_2185012 |
| Mouse anti-α-tubulin (DM1α) | Sigma | Cat#T6199; RRID:AB_477583 |
| Rabbit anti-Septin 2 (clone: a5N5) | Kind gift form M. Kinoshita (Nagoya University, Japan) | N/A |
| Rabbit anti-Septin 6 | Kind gift form M. Kinoshita (Nagoya University, Japan) | N/A |
| Rabbit anti-Septin 7 | IBL America | Cat#18991; RRID:AB_1630825 |
| Rabbit anti-CHMP4B | PTG labs | Cat# 15669-1-AP |
| Mouse anti-TSG101 (clone C-52) | Santa Cruz | Cat# sc-7964, RRID:AB_671392 |
| Mouse anti-ALIX | Santa Cruz | Cat# sc-53540, RRID:AB_673819 |
| Rabbit anti-VPS25 | Abclonal | Cat# A13132, RRID:AB_2759982 |
| Rabbit anti-VPS22 | Abclonal | Cat# A10583, RRID:AB_2758124 |
| Rabbit anti-CHMP2B | Abclonal | Cat# A13410, RRID:AB_2760272 |
| Rabbit anti-TSG101 | PTG labs | Cat# 14497-1-AP, RRID:AB_2208090 |
| Rabbit anti-CHMP6 | PTG labs | Cat# 16278-1-AP, RRID:AB_2079498 |
| Rabbit anti-VPS25 | PTG labs | Cat# 15669-1-AP, RRID:AB_2215019 |
| Mouse anti-mCherry | Abcam | Cat# |
| Mouse anti-His tag | Qiagen | Cat# Cat# 34440, RRID:AB_2714179 |
| Rabbit anti-CHMP4B | Santa Cruz | Cat# sc-134946, RRID:AB_10608470 |
| Rabbit anti-GST | Santa cruz | Cat# sc-459, RRID:AB_631586 |
| goat-anti rabbit IRDye® 680RD | LI-COR | Cat# 925-68071; RRID:AB_2721181 |
| goat anti-mouse IRDye® 800RD | LI-COR | Cat# 925-32210; RRID:AB_2687825 |
| Bacterial and Virus Strains | ||
| Escherichia coli BL21 (DE3) | Invitrogen | Cat#C600003 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Purified 6XHis-hSEPT9 (GenBank accession number NP_001106963; isoform a ) protein | [44] | N/A |
| Purified 6XHis-hSEPT9 Δ(PTAP)2x | This paper | N/A |
| GST-TSG101-UEV | This paper | N/A |
| Ni-NTA agarose beads | Macherey-Nagel | Cat# 745400.25 |
| Glutathione agarose beads | Thermo Scientific | Cat# 16100 |
| RFP-Trap®_A beads | ChromoTek | Cat# rta-10 |
| Critical Commercial Assays | ||
| Quick Change II site-mutagenesis kit | Agilent Technologies | Cat#200523 |
| Duolink® PLA – Protein Detection Technology | Sigma - Aldrich | Cat# DUO92101-1KT |
| Zenon™Alexa Fluor™ 568 Rabbit IgG Labeling Kit |
ThermoFisher Scientific | Cat# Z-25306; RRID:AB_2736952 |
| Experimental Models: Cell Lines | ||
| Madin Darby kidney canine epithelia cells (MDCK) II/G | Kind gift from J. Nelson, Stanford University, CA | ECACC #00062107 |
| MDCK mCherry- SEPT9 | [42] | N/A |
| MDCK mCherry- SEPT2 | [13] | N/A |
| MDCK SEPT2-YFP | [12] | N/A |
| MDCK GFP-SEPT6 | This paper | N/A |
| MDCK GFP-CHMP4B | This paper | N/A |
| Oligonucleotides – see Table S1 for sequences of primers used for this study | ||
| Recombinant DNA | ||
| GFP-TSG101 | Kind gift from Dr. Paul Bieniasz (The Rockefeller University, NY) | N/A |
| GFP-CHMP4B | Kind gift from Dr. Paul Bieniasz, The Rockefeller University, NY | N/A |
| GFP-CHMP2A | Addgene | Cat# 31805 , deposited by the Daniel Gerlich Lab |
| CHMP6-GFP | Addgene | Cat #31806 , deposited by the Daniel Gerlich Lab |
| GFP-VPS25 |
Kind gift from Dr. Natalie Elia, Ben Gurion University of Negev, Israel [20] | N/A |
| GFP-VPS36 | Kind gift from Dr. Natalie Elia, Ben Gurion University of Negev, Israel [20] | N/A |
| GFP-VPS22 | Kind gift from Dr. Natalie Elia, Ben Gurion University of Negev, Israel [20] | N/A |
| pEGFP-C1-VPS4 |
Kind gift from Dr. Wes Sundquist, University of Utah School of Medicine, UT | N/A |
| pEGFP-C1-α-tubulin | Kind gift from J. Nelson, Stanford University, CA [13] | N/A |
| pBiFC-VN173 | Addgene | Cat#22010 deposited by the Chang-Deng Hu lab |
| pBiFC-VC155 | Addgene | Cat#22010 deposited by the Chang-Deng Hu lab |
| SEPT9_VN | This paper | N/A |
| TSG101_VC | This paper | N/A |
| TSG101_VN | This paper | N/A |
| pmCherry-C1-SEPT2 | [13] | N/A |
| pmCherry -C1-SEPT9 | [42] | N/A |
| pYFP-N1- (mouse) SEPT2 | [12] | N/A |
| pEGFP-C3-SEPT6 | [43] | N/A |
| pmCherry-N-hSEPT9 | [42] | N/A |
| pGFP-VR-S-canine S9 shRNA-rescuehSEPT9-FL-mCherry | This paper | N/A |
| pGFP-VR-S-canine S9 shRNA1-rescuehSEPT9-Δ(PTAP)2x -mCherry | This paper | N/A |
| GST-TSG101 UEV | This paper | N/A |
| His-hSEPT9 | [44] | N/A |
| His-hSEPT9- Δ(PTAP)2x | This paper | N/A |
| Software and Algorithms | ||
| Slidebook | 3i | https://www.intelligent-imaging.com/ ; RRID:SCR_014300 |
| Fiji (ImageJ) | NIH | https://imagej.nih.gov/ij/ ; RRID:SCR_002285 |
| GraphPad Prism | GraphPad | https://www.graphpad.com/scientific-software/prism/ ; RRID:SCR_002798 |
| MATLAB | MATLAB | http://www.mathworks.com/products/matlab/; RRID:SCR_001622 |