Table 1.

Key resources table

Reagent or resourceSourceIdentifier
Antibodies 
Rabbit polyclonal anti-MYCT1 Abcam Cat# ab139945, RRID:AB_2861408 
Mouse monoclonal anti-IFITM2 Thermo Fisher Scientific Cat# CF811628 
Rabbit polyclonal anti-IFITM3 Proteintech Cat# 11714-1-AP, RRID:AB_2295684 
Goat polyclonal anti VE-cadherin R&D Systems Cat# AF1002, RRID:AB_2077789 
Mouse anti-β-catenin BD Biosciences Cat# 610154, RRID:AB_397555 
Goat polyclonal anti-VEGFR2 R&D Systems Cat# AF644, RRID:AB_355500 
Rabbit polyclonal anti-β-catenin Sigma-Aldrich Cat# 06-734, RRID:AB_310231 
Rabbit polyclonal anti-GFP Abcam Cat# ab290, RRID:AB_303395 
Mouse monoclonal anti-Rab5 Cell Signaling Technology Cat# 46449, RRID:AB_2799303 
Rabbit monoclonal anti-Rab5 Cell Signaling Technology Cat# 3547, RRID:AB_2300649 
Rabbit monoclonal anti-Rab7 Abcam Cat# ab137029, RRID:AB_2629474 
Mouse monoclonal anti-Lamp1 Santa Cruz Biotechnology Cat# sc-20011, RRID:AB_626853 
Rabbit monoclonal anti-phospho-S6 ribosomal protein (Ser240/244) Cell Signaling Technology Cat# 5364, RRID:AB_10694233 
Mouse monoclonal anti-S6 ribosomal protein Cell Signaling Technology Cat# 2317, RRID:AB_2238583 
Rabbit anti-p70/S6K (Thr389) Cell Signaling Technology Cat# 9234, RRID:AB_2269803 
Rabbit anti-p70/S6K Cell Signaling Technology Cat# 9202, RRID: AB_2943679 
Rabbit polyclonal anti-Mat2a Proteintech Cat No. 55309-1-AP, RRID:AB_2881303 
Rabbit monoclonal anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Cell Signaling Technology Cat# 4376, RRID:AB_331772 
Rabbit monoclonal anti-p44/42 MAPK (Erk1/2) Cell Signaling Technology Cat# 4695, RRID:AB_390779 
Rabbit monoclonal anti-phospho-Akt (Thr308) Cell Signaling Technology Cat# 13038, RRID:AB_2629447 
Rabbit monoclonal anti-phospho-Akt (Ser473) Cell Signaling Technology Cat# 4060, RRID:AB_2315049 
Rabbit polyclonal anti-AKT Cell Signaling Technology Cat# 9272, RRID:AB_329827 
Rabbit polyclonal anti-GAPDH Sigma-Aldrich Cat# G9545, RRID:AB_796208 
Mouse monoclonal anti-Vinculin Sigma-Aldrich Cat# V9131, RRID:AB_477629 
Mouse monoclonal anti-Pecam1 Thermo Fisher Scientific Cat# MA5-13188, RRID:AB_10982120 
Rat anti-Pecam1 BD Biosciences Cat# 553370, RRID:AB_394816 
Rat monoclonal anti-endomucin Thermo Fisher Scientific Cat# 14-5851-81, RRID:AB_891529 
Rabbit polyclonal anti-Pecam1 Abcam Cat# ab28364, RRID:AB_726362 
Rabbit monoclonal anti-Erg Abcam Cat# ab92513, RRID:AB_2630401 
Goat polyclonal anti-FABP4 R&D Systems Cat# AF1443, RRID:AB_2102444 
Rabbit monoclonal anti-Perilipin Cell Signaling Technology Cat# 9349, RRID:AB_10829911 
Rat monoclonal anti-Pecam1 (BV421 conjugated) BioLegend Cat# 102423, RRID:AB_2562186 
Rabbit anti-H3K27ac Abcam Cat# ab4729, RRID:AB_2118291 
Rabbit anti-GAPDH Sigma-Aldrich Cat# G9545, RRID:AB_796208 
Mouse anti-Vimentin Dako Cat# M0725, RRID:AB_10013485 
CD45-APC/Cy7, clone 30-F11 BioLegend Cat# 561037, RRID:AB_10563075 
CD45-PE, clone 30-F11 BioLegend Cat# 103105, RRID:AB_312970 
CD31-BV421, clone 390 BD Pharmingen Cat# 102423, RRID:AB_2562186 
CD31-PE, clone 390 eBioscience Cat# 12-0311-81 
Sca1-PE/Cy7, clone D7 BioLegend Cat# 108113, RRID:AB_493597 
Bacterial and virus strains 
Ad-MYCT1 and Ad-GFP adenoviruses This study ​ 
Biological samples 
Human brain samples Hôpitaux universitaires de Genève (HUG) (Geneva, Switzerland) CCER 2022-02109 
Human fat samples Lausanne University Hospital (CHUV) (Lausanne, Switzerland) CER-VD 38/15, PB_2016-01185 
Human normal and cancer samples CHUV (Lausanne, Switzerland) CER-VD 464/11 
Chemicals, peptides, and recombinant proteins 
Tamoxifen Sigma-Aldrich T5648 
Cremophor EL Sigma-Aldrich C5135 
PFA Sigma-Aldrich 158127 
EBM2 medium Lonza CC-3156 
EGM2 SingleQuots Lonza CC-4176 
Opti-MEM medium Gibco 51985-026 
PBS containing MgCl2 and CaCl2 Sigma-Aldrich D8662 
Lipofectamine RNAiMAX Thermo Fisher Scientific 13778-075 
OPP Jena Bioscience NU-931-05 
Vitamin solution Sigma-Aldrich R7256 
Amino acid solution Sigma-Aldrich R7131 
GlutaMAX Thermo Fisher Scientific 35050038 
Rapamycin LC Laboratories R-5000 
PhosSTOP Roche 04906837001 
Complete mini, EDTA-free Protease inhibitor Roche 11836170001 
Histodenz Sigma-Aldrich D2158 
Liberase TL Sigma-Aldrich 05401020001 
HBSS Gibco 14175-053 
Red blood cell lysis Thermo Fisher Scientific 00-4333-57 
Fibronectin Roche 10838039001 
Dynasore Sigma-Aldrich 324410 
Cycloheximide Enzo Life Sciences ALX-380-269 
Fluorescein isothiocyanate–dextran (10 kDa) Sigma-Aldrich FD10S 
DQ Green BSA Thermo Fisher Scientific D12050 
Evans blue Sigma-Aldrich E2129 
DMEM Sigma-Aldrich D5030 
Insulin Sigma-Aldrich I9278 
Dexamethasone Sigma-Aldrich D4902 
3-Isobutyl-1-methylxanthine (IBMX) Enzo Life Sciences BML-PD140 
Indomethacin Enzo Life Sciences ALX-270-086 
Recombinant PNGase F Provided by P. Schneider (Department of Immunobiology [DIB], University of Lausanne [Unil], Lausanne, Switzerland) 704S; NEB 
Fluoromount-G mounting medium Thermo Fisher Scientific 00-4959-52 
Aquatex Merck 1085620050 
Heparin Sigma-Aldrich H3149 
Tamoxifen diet Harlan TD.55125 
HFD Research Diet D12492 
Control diet Research Diet D12450B 
Anti-rat IgG magnetic Dynabeads Thermo Fisher Scientific 11035 
7-AAD eBioscience 00-6993-50 
Critical commercial assays 
Duolink in situ Detection Reagents Orange Sigma-Aldrich DUO92007 
Duolink in situ PLA probe anti-mouse PLUS Sigma-Aldrich DUO92001 
Duolink in situ PLA probe anti-rabbit MINUS Sigma-Aldrich DUO92005 
Duolink in situ PLA probe anti-goat PLUS Sigma-Aldrich DUO92003 
RNeasy Plus mini kit Qiagen 74134 
Transcriptor First Strand cDNA Synthesis kit Roche 04 896 866 001 
SensiFast Sybr Lo-Rox Mix Bioline BIO-94020 
SuperSignal West Femto Maximum Sensitivity Substrate Thermo Fisher Scientific 34095 
Pierce BCA Protein assay kit Thermo Fisher Scientific 23227 
Lipofectamine RNAiMAX Thermo Fisher Scientific 3778075 
Protein G magnetic Dynabeads Thermo Fisher Scientific 10003D 
Tyramide SuperBoost Kits with Alexa Fluor Tyramides Thermo Fisher Scientific B40931 
Deposited data 
GEO accession number GSE294341 This study https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE294341 
scRNA-seq data of human ECs from several organs (Barnett et al., 2024) https://www.vascularcellatlas.org/ 
Experimental models: Cell lines 
Primary HIECs Isolated in the lab Anonymous donors, CHUV, Lausanne 
Primary human pulmonary and adipose tissue microvascular ECs Innoprot, PromoCell P10552, P10781 C-12281 
Primary HUVECs Lonza C2519A 
Human embryonic kidney 293T cells ATCC CRL-1573 
SW480 cells ATCC CCL-228 
LLC cells ATCC CRL-1642 
Experimental models: Organisms/strains 
Myct1flox/+ mouse This study InGenious Targeting Laboratory Inc. 
Pdgfb-CreERT2 mouse (Claxton et al., 2008) Provided by M. Fruttiger (UCL, London, UK) 
Tsc1flox/+ mouse (Uhlmann et al., 2002) Provided by O. Dormond (UNIL, Lausanne, Switzerland) 
Oligonucleotides 
Primers qPCR 
18 s (human/mouse) forward Eurofins 5′-AGG​AAT​TCC​CAG​TAA​GTG​CG-3′ 
18 s (human/mouse) reverse Eurofins 5′-GCC​TCA​CTA​AAC​CAT​CCA​A-3′ 
MYCT1 (human/mouse) forward Eurofins 5′-CTT​TCC​ATC​CCT​TTC​TGC​AA-3′ 
MYCT1 (human/mouse) reverse Eurofins 5′-GTG​CTG​ATG​GTG​GGA​GAG​AT-3′ 
IFITM2 (human) forward Eurofins 5′-GTC​ACC​ATG​AAC​CAC​ATT​GTG​CAA​AC-3′ 
IFITM2 (human) reverse Eurofins 5′-CCC​CCA​GCA​TAG​CCA​CTT​CC-3′ 
IFITM3 (human) forward Eurofins 5′-ACC​ATG​AAT​CAC​ACT​GTC​CAA​ACC​TT-3′ 
IFITM3 (human) reverse Eurofins 5′-CCA​GCA​CAG​CCA​CCT​CG-3′ 
Pecam1 (mouse) forward Eurofins 5′-AAC​AGA​AAC​CCG​TGG​AGA​TG-3′ 
Pecam1 (mouse) reverse Eurofins 5′-GTC​TCT​GTG​GCT​CTC​GTT​CC-3′ 
Cdh5 (mouse) forward Eurofins 5′-CCC​ACG​AAG​TCC​CTG​GAC​TAT​G-3′ 
Cdh5 (mouse) reverse Eurofins 5′-GGT​CTG​TGG​CCT​CAA​TGT​AGA​ATG-3′ 
Ucp1 (mouse) forward Eurofins 5′-CTG​CCA​GGA​CAG​TAC​CCA​AG-3′ 
Ucp1 (mouse) reverse Eurofins 5′-GCC​ACA​AAC​CCT​TTG​AAA​AA-3′ 
Cpt1b (mouse) forward Eurofins 5′-CCC​ATG​TGC​TCC​TAC​CAG​AT-3′ 
Cpt1b (mouse) reverse Eurofins 5′-CCT​TGA​AGA​AGC​GAC​CTT​TG-3′ 
Prdm16 (mouse) forward Eurofins 5′-CAG​CAC​GGT​GAA​GCC​ATT​C-3′ 
Prdm16 (mouse) reverse Eurofins 5′-GCG​TGC​ATC​CGC​TTG​TG-3′ 
Cebpb (mouse) forward Eurofins 5′-CGC​AAC​CTG​GAG​ACG​CAG​CA-3′ 
Cebpb (mouse) reverse Eurofins 5′-GGC​TCG​GGC​AGC​TGC​TTG​AA-3′ 
Ppara (mouse) forward Eurofins 5′-GAC​AAG​GCC​TCA​GGG​TAC​CA-3′ 
Ppara (mouse) reverse Eurofins 5′-GCC​GAA​TAG​TTC​GCC​GAA​A-3′ 
siRNA 
All Stars negative control Qiagen 1027281 
MYCT1 (human) #6 (A) Qiagen SI04203052 
MYCT1 (human) #8 (B) Qiagen SI04281760 
IFITM2/3 (human) #5 Qiagen SI04179721 
RAB5A (human) #10 Qiagen SI03111115 
Software and algorithms 
GraphPad Prism 10 GraphPad Software https://www.graphpad.com/ 
Fiji version 1.54f (Schindelin et al., 2012) https://fiji.sc 
CellProfiler version 4.2.4 CellProfiler https://cellprofiler.org 
Affinity Photo 2 Affinity https://affinity.serif.com 
Affinity Publisher 2 Affinity https://affinity.serif.com 
Affinity Designer 2 Affinity https://affinity.serif.com 
BioRender BioRender https://biorender.com 
FlowJo version 10 BD Biosciences https://www.flowjo.com/ 
Cellpose version 2.1.1 (Stringer et al., 2021) https://github.com/mouseland/cellpose 
Napari version 0.4.17 (Sofroniew et al., 2025) https://napari.org/0.4.17/index.html 
Scikit-image version 0.19.3 (van der Walt et al., 2014) https://scikit-image.org/ 
Python version 3.9.7 Python Software Foundation https://www.python.org/ 
PantherDB version 19.0 PantherDB https://pantherdb.org/ 
Cell Ranger version 6.0.0 10x Genomics https://www.10xgenomics.com/support/software/cell-ranger/downloads 
R versions 4.3.1 and 4.1.2 The R Project for Statistical Computing https://www.r-project.org/ 
Seurat package versions 5.0.1 and 4.3.0 (Hao et al., 2024) https://cran.r-project.org/web/packages/Seurat/index.html 
clusterProfiler package version 4.2.2 (Xu et al., 2024) https://bioconductor.org/packages/release/bioc/html/clusterProfiler.html 
msigdbr package version 7.5.1 (Dolgalev, 2025) https://cran.r-project.org/web/packages/msigdbr/index.html 
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