Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | anti-PTRF (Rabbit polyclonal) |
Abcam | Cat # ab48824 RRID:AB_88224 |
IF(1:100) |
| Antibody | anti-EHD2 (Rabbit polyclonal) |
(Morén et al., 2012) | RRID:AB_2833022 | IF (1:500), WB(1:2000) |
| Antibody | anti-clathrin heavy chain (clone 23; mouse monoclonal) | BD Transduction Laboratories | Cat # 610499 RRID:AB_397865 |
WB (1:1000) |
| Antibody | anti-caveolin-1 (Rabbit, polyclonal) |
Abcam | Cat # ab2910 RRID:AB_303405 |
IF (1:500) WB (1:10 000) |
| Antibody | anti-rabbit IgG secondary antibody coupled to Alexa Fluor 647 (Goat polyclonal) | Thermo Fisher Scientific | Cat # A21246 RRID:AB_2535814 |
IF (1:300) |
| Chemical compound, drug | 1,2-dioleoyl-sn -glycero-3-phosphoethanolamine (DOPE) |
Avanti Polar Lipids | Cat # 850725 | |
| Chemical compound, drug | 1,2-dioleoyl-3-trimethylammonium-propane (chloride salt) (DOTAP) | Avanti Polar Lipids | Cat # 890890 | |
| Chemical compound, drug | Bodipy FL C5- Ganglioside GM1 | Thermo Fisher Scientific | Cat # B13950 | |
| Chemical compound, drug | Bodipy FL C5-LacCer | This study (Gretskaya and Bezuglov, 2013) |
||
| Chemical compound, drug | Bodipy FL C5-ceramide | Thermo Fisher Scientific | Cat # D3521 | |
| Chemical compound, drug | Bodipy-Cholesterol | Avanti Polar Lipids | Cat # 810255 | |
| Chemical compound, drug | Bodipy FL C12-spinghomyelin | Thermo Fisher Scientific | Cat # D7711 | |
| Chemical compound, drug | Bodipy FL C5-spinghomyelin | Thermo Fisher Scientific | Cat # D3522 | |
| Chemical compound, drug | Bodipy FL succinimidyl ester (EverFluor FL) | Setareh Biotech | Cat # 7150 | |
| Chemical compound, drug | D-lactosyl-ß1–1'-D-erythro-sphingosine | Avanti Polar Lipids | Cat # 860542 | |
| Chemical compound, drug | D-lactosyl-ß−1,1' N-palmitoyl-D-erythro-sphingosine | Avanti Polar Lipids | Cat # 860576 | |
| Chemical compound, drug | Cholesterol | Sigma Aldrich | Cat # C3045 | |
| Chemical compound, drug | N-Palmitoyl-D-sphingomyelin | Sigma Aldrich | Cat# 91553 | |
| Chemical compound, drug | Sphingomyelinase (SMase) (from Bacillus cereus) | Sigma Aldrich | Cat# S7651 | |
| Chemical compound, drug | Myriocin (from Mycelia sterilia) | Sigma Aldrich | Cat# M1177 | |
| Chemical compound, drug |
Paraformaldehyde 16% | Electron Microscopy Sciences | Cat # 15710 | |
| Chemical compound, drug | Paraformaldehyde | Thermo Fisher Scientific | Cat # PA0095 | |
| Chemical compound, drug | Glutaraldehyde 25% | Taab Laboratory Equipment Ltd | Cat # G011/2 | |
| Chemical compound, drug | 3-isobutyl-1-methylxanthine (IBMX) | Sigma Aldrich | Cat # I5879 | |
| Chemical compound, drug |
Dexamethasone (Dex) | Sigma Aldrich | Cat # D4902 | |
| Chemical compound, drug | Insulin | Sigma Aldrich | Cat # I0516 | |
| Chemical compound, drug | rosiglitazone | Cayman Chemicals | 71740 | |
| Chemical compound, drug | Lipofectamine2000 | Thermo Fisher Scientific | Cat # 11668019 | |
| Chemical compound, drug | N,N-Diisopropylethylamine | Sigma Aldrich | Cat # D125806 | |
| Chemical compound, drug | N,N-Dimethylformamide, anhydrous | Sigma Aldrich | Cat # 227056 | |
| Chemical compound, drug | Chloroform | VWR | Cat # VWRC22711.324 | |
| Chemical compound, drug | Methanol | Thermo Fisher Scientific | Cat # 15394528 | |
| Chemical compound, drug |
Doxycycline hyclate | Sigma Aldrich | Cat # D9891 | |
| Chemical compound, drug | Hygromycin B | Thermo Fisher Scientific | Cat # 10687010 | |
| Chemical compound, drug | Blasticidin S HCl | Thermo Fisher Scientific | Cat # R210-01 | |
| Cell line (Homo-sapiens) | HeLa (cervix) |
ATCC | Cat # CRM-CCL-2 RRID:CVCL_0030 |
|
| Cell line (M. musculus) | 3T3-L1 (Pre-adipocytes) | ATCC | Cat # ATC-CL-173 RRID:CVCL_0123 |
|
| Cell line (Homo-sapiens) | HeLa FlpIn T-Rex Caveolin1-mCherry | This study | ||
| Cell line (Homo-sapiens) | HeLa FlpIn T-Rex EHD2pTagBFP-P2A-Caveolin1-mCherry | This study | ||
| Transfected construct (human) | siRNA to EHD2 (Stealth) |
Thermo Fisher Scientific | Cat#1299001 Assay ID HSS121265 | |
| Transfected construct (human) | Scrambled 353 siRNA (Stealth) |
Thermo Fisher Scientific | Cat# 12935300 | |
| Transfected construct (human) | siRNA to Caveolin1 (Stealth) |
Thermo Fisher Scientific | Cat #1299001 Assay ID HSS141467 | |
| Recombinant DNA reagent | Rab5-TagBFP | (Francis et al., 2015) | under control of PCMV IEpromotor in pTagBFP-N vector | |
| Recombinant DNA reagent | EHD2-TagBFP | This study | under control of PCMV IEpromotor in pTagBFP-N vector | |
| Recombinant DNA reagent | EHD2 I157Q-TagBFP | This study | under control of PCMV IEpromotor in pTagBFP-N vector | |
| Recombinant DNA reagent | EHD2-mCherry | (Hoernke et al., 2017) | under control of PCMV IEpromotor in pmCherry-N1 vector | |
| Recombinant DNA reagent | mCherry-EHD2 | This study | under control of PCMV IEpromotor in pmCherry-C1 vector | |
| Recombinant DNA reagent | EHD2 I157Q-mCherry | This study | under control of PCMV IEpromotor in pmCherry-N1 vector | |
| Recombinant DNA reagent | mCherry-EHD2 I157Q | This study | under control of PCMV IEpromotor in pmCherry-C1 vector | |
| Recombinant DNA reagent | pCDNA/FRT/TO caveolin1-mCherry | This study | under control of PCMV IE promotor | |
| Recombinant DNA reagent | pCDNA/FRT/TO EHD2-pTagBFP-P2A-caveolin1-mCherry | This study | under control of PCMV IE promotor | |
| Software, algorithm | ImageJ/Fiji | Fiji (Schindelin et al., 2012) |
RRID:SCR_002285 | http://fiji.sc/ |
| Software, algorithm | Imaris x64 9.1.2 | Bitplane | RRID:SCR_007370 | http://www.bitplane.com/imaris |
| Software, algorithm | Prism 5.0 | GraphPad | RRID:SCR_002798 | https://www.graphpad.com/scientific-software/prism/ |
| Software, algorithm | Photoshop CS6 | Adobe | RRID:SCR_014199 | https://www.adobe.com/se/products/photoshop.html |
| Software, algorithm | Illustrator CS6 | Adobe | RRID:SCR_010279 | https://www.adobe.com/se/products/photoshop.html |
| Software, algorithm | Maps 3.3 | FEI | https://www.fei.com/software/maps/ | |
| Software, algorithm | Zen interface 2.3 | Zeiss | RRID:SCR_013672 | https://www.zeiss.com/microscopy/int/products/microscope-software/zen.html |
| Software, algorithm | Nis Elements 4.3 | Nikon | RRID:SCR_014329 | https://www.nikoninstruments.com/en_EU/Products/Software |
| Other | Aluminum backed silica gel plates (median pore size 60 Å, fluorescent indicator 254 nm) | Fisher Scientific | Cat # 10517771 | |
| Other | Chromatography grade silica gel (0.035–0.070 mm, 60 Å) | Acros Organic | Cat # 240360050 | |
| Other | Formvar TEM grids | Taab Laboratory Equipment Ltd | Cat # F005 | |
| Other | CS-25R17 coverlips (TIRF) | Warner Instruments | Cat # 64–0735 | |
| Other | CS-25R15 coverlips | Warner Instruments | Cat # 64–0715 | |
| Other | Precision cover glasses thickness No. 1.5H | Paul Marienfeld GmbH and Co. KG | Cat # 0117520 | |
| Other | PD-10 columns | GE Healthcare | Cat # 17-0851-0 | |
| Other | MatTek dishes (35 mm dish, high tolerance 1.5) | MatTek Corporation | Cat # P35G-0.170–14 C | |
| Other | HCS LipidTOX Deep Red Neutral Lipid Stain | Thermo Fisher Scientific | Cat # H34477 | |
| Other | Dulbecco's Modified Eagle Medium (DMEM) | Thermo Fisher Scientific | Cat # 41966052 | |
| Other | Opti-MEM I Reduced Serum Medium | Thermo Fisher Scientific | Cat # 31985070 | |
| Other | Dulbecco's Modified Eagle Medium (DMEM), no phenol red | Thermo Fisher Scientific | Cat # 21063029 | |
| Other | Sodium pyruvate | Thermo Fisher Scientific | Cat # 11360039 | |
| Other | Dako Fluorescent Medium | Dako | Cat # S3023 | |
| Other | Alexa Fluor 647 C2 Maleimide | Thermo Fisher Scientific | Cat # A20347 | |
| Other | DAPI (4',6-Diamidino-2-Phenylindole, Dilactate) | Thermo Fisher Scientific | Cat # D3571 | |
| Other | Fetal bovine serum | Thermo Fisher Scientific | Cat # 16000044 |