| Antibodies |
| Mouse monoclonal anti-NS1 (7E11) in house conjugated to Alexa 568 |
Gift from R. Putnak (WRA) in-house conjugated |
NA |
| Mouse monoclonal IgG anti-His Tag (AD1.1.10’)(AlexaFluor®647) |
NOVUS Biologicals |
Cat# NB100–64768AF647 |
| Mouse monoclonal IgG anti-human CD138 (Syndecan-1) purified. Clone: DL-101 |
eBioscience |
Cat#14–1389–82; RRID:AB_467489 |
| Rabbit polyclonal IgG anti-heparanase 1 (HPA1)(H-80) |
Santa Cruz Biotechnology |
Cat# sc-25825; RRID:AB_2120854 |
| Mouse monoclonal IgG anti-human Cathepsin L Purified. |
Invitrogen |
Cat# BMS166; RRID:AB_10597594 |
| Rabbit polyclonal IgG anti-neuraminidase 1 (Neu1)(H-300) |
Santa Cruz Biotechnology |
Cat# sc-32936; RRID:AB_2298197 |
| Mouse monoclonal IgG anti-neuraminidase 1 (Neu1)(F-8) |
Santa Cruz Biotechnology |
Cat# sc-166824; RRID:AB_2149061 |
| Goat polyclonal IgG anti-human ganglioside sialidase (Neu3)(N-18) |
Santa Cruz Biotechnology |
Cat# sc-55826; RRID:AB_2149075 |
| Rabbit polyclonal IgG anti-human neuraminidase 2 (Neu2) |
Thermo Scientific (Invitrogen™) |
Cat# PA5–35114; RRID:AB_2552424 |
| Rabbit polyclonal IgG anti-human neuraminidase 3 (Neu3) |
Thermo Scientific (Invitrogen™) |
Cat# PA5–45518; RRID:AB_2576484 |
| Rabbit polyclonal IgG anti-human GAPDH (FL-335) |
Santa Cruz Biotechnology |
Cat# sc-25778; RRID:AB_10167668 |
| Mouse IgM anti-heparan sulfate (F58–10E4) |
Amsbio |
Cat# 370255–1; RRID:AB_10891554 |
| Mouse monoclonal IgG anti-6x His epitope Tag (HIS.H8) |
Thermo Scientific (Invitrogen™) |
Cat# MA1–21315; RRID:AB_557403 |
| Rabbit polyclonal anti-Alpha tubulin (loading control) |
Abcam |
Cat# Ab4074; RRID:AB_2288001 |
| Rabbit polyclonal anti-Rab5 (early endosome marker) |
Abcam |
Cat# Ab18211; RRID:AB_470264 |
| Goat anti-mouse IgG (AlexaFluor®647) |
Abcam |
Cat# ab150115; RRID:AB_2687948 |
| Donkey anti-mouse IgG (AlexaFluor®647) |
Thermo Scientific (Invitrogen™) |
Cat# A31571; RRID:AB_162542 |
| Goat anti-rabbit IgG (AlexaFluor®568) |
Abcam |
Cat# ab175471; RRID: AB_2576207 |
| Donkey anti-rabbit IgG (AlexaFluor®568) |
Abcam |
Cat# ab175470 |
| Donkey anti-rabbit IgG (AlexaFluor®647) |
Abcam |
Cat# ab150075; RRID: AB_2752244 |
| Donkey anti-goat IgG (AlexaFluor®568) |
Abcam |
Cat# ab175474; RRID:AB_2636995 |
| Donkey anti-goat IgG (AlexaFluor®647) |
Abcam |
Cat# ab150131; RRID:AB_2732857 |
| Donkey anti-mouse IgM (AlexaFluor®488) |
Jackson Immuno Research |
Cat# 715–545–140; RRID:AB_2340845 |
| Goat anti-mouse IgG (AlexaFluor®680) |
Thermo Scientific (Invitrogen™) |
Cat# A21057; RRID:AB_141436 |
| Goat anti-mouse IgG (AlexaFluor®750) |
Thermo Scientific (Invitrogen™) |
Cat# A21037; RRID:AB_1500644 |
| Donkey anti-mouse IgG (AlexaFluor®680) |
Abcam |
Cat# ab175774 |
| Donkey anti-mouse IgG (AlexaFluor®750) |
Abcam |
Cat# ab175738 |
| Goat anti-rabbit IgG (AlexaFluor®680) |
Thermo Scientific (Invitrogen™) |
Cat# A21076; RRID:AB_141386 |
| Goat anti-rabbit IgG (AlexaFluor®750) |
Thermo Scientific (Invitrogen™) |
Cat# A21039; RRID: AB_1500687 |
| Donkey anti-rabbit IgG (AlexaFluor®680) |
Abcam |
Cat# ab186692; RRID:AB_2725787 |
| Donkey anti-rabbit IgG (AlexaFluor®750) |
Abcam |
Cat# ab175728 |
| Donkey anti-human IgG (AlexaFluor®680) |
Jackson Immuno Research |
Cat# 709–625–149; RRID:AB_2340582 |
| Goat anti-mouse IgG (AlexaFluor®488) |
Abcam |
Cat# Ab150117; RRID:AB_2688012 |
| Chemicals, Peptides, and Recombinant Proteins |
| Gelatin solution, Type B, 2% in H2O, tissue culture grade |
Sigma |
Cat# G1393–20ML |
| Wheat germ agglutinin (AlexaFluor®647) |
Thermo Scientific (Invitrogen™) |
Cat# W32466
|
| Dextran, Alexa Fluor® 680; 10,000 MW, Anionic, Fixable, 5 m |
Life Technologies |
Cat# D34680
|
| Formamide |
VWR Life science |
Cat# 0314 |
| Fluorescein isothiocyanate (FITC)–dextran, average mol wt 70,000 |
Sigma |
Ca# FD70S-100MG |
| Recombinant dengue virus 2 NS1 (strain 16681) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant Zika virus NS1 (Uganda MR766) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant Zika virus NS1 (Suriname Z1106033) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant West Nile virus NS1 (NY99) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant Japanese encephalitis virus NS1 (SA-14) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant Yellow fever virus NS1 (17D) protein produced in 293 human cells. C-terminal 6x His-Tag |
The Native Antigen Company |
NA |
| Recombinant Dengue 2 Envelope (E) protein (DENV 2 E80%) produced in Sf21 cells. |
In-house produced (gift from Aravinda M. de Silva) |
NA |
| Recombinant DENV2/WNV Chimera (aa101–135WNV) produced in 293 human cells. C-terminal 6x His-Tag |
In-house produced (this paper) |
NA |
| Human TNF-alpha Recombinant Protein |
Affymetrix/eBioscience |
Cat# 14–8329–63 |
| Vascular endothelial growth factor (VEGF) |
Sigma |
Cat# V7259–10UG |
| Critical Commercial Assays and kits |
| MR-(FR)2 Magic Red® Cathepsin-L substrate |
Inmunochemistry |
Cat# 6138 |
| Corning Transwell Multiple Well Plate with Permeable Polycarbonate Membrane Inserts |
Fisher Scientific (Corning) |
Cat# 07–200–147 |
| Corning® Transwell®-COL collagen-coated membrane inserts |
Sigma (Corning) |
Cat# CLS3495–24EA |
| Endpoint Chromogenic LAL QCL-1000 120 Test Kit |
Lonza |
Cat# 50–647U |
| EBMTM-2 Basal Medium |
Lonza |
Cat#: CC-3156 |
| EGMTM-2 MV Microvascular Endothelial SingleQuotsTM Kit |
Lonza |
Cat#: CC-4147 |
| Endothelial Cell Medium |
ScienCell |
Cat#: 1001 |
| Endothelial Cell Medium |
Cell Biologics |
Cat# 1166 |
| Endothelial Cell Growth Medium 2 (Ready-to-use) |
PromoCell |
Cat# C-22011 |
| FreeStyle serum free medium |
Thermo Scientific |
Cat# 12338018 |
| Alexa Fluor 568 Antibody Labeling Kit |
Thermo Scientific (Invitrogen) |
Cat# A20184 |
| Experimental Models: Cell Lines |
| Human pulmonary microvascular endothelial cell line (HPMEC). Clone HPMEC.ST1–6R |
Gift from J.C. Kirkpatrick, Johannes-Gutenberg University |
NA |
| Human umbilical vein cord endothelial cells (HUVEC) |
Gift from Melisa Lodoen, UC Irvine (Manufacturer: Lonza) |
Cat#:C2517A |
| Human dermal microvascular endothelial cell line (HMEC-1) |
Gift from Matthew Welch, UC Berkeley |
NA |
| Human brain microvascular endothelial cells (HBMEC) |
Gift from Ana Rodriguez, NYU (Manufacturer: ScienCell) |
Cat#: 1000 |
| Human liver sinusoidal microvascular endothelial cells (HLSEC) |
Cell Biologics |
Cat# H-6017 |
| Human 293F cells |
UC Berkeley Cell Culture facility |
Custom order |
| Experimental Models: Organisms/Strains |
|
|
| Mouse: C57BL/6J |
The Jackson Laboratory |
Stock No: 000664 |
| Oligonucleotides |
| Primer oligonucleotides for Mycoplasma screening sense GPO-3 5′GGGAGCAAACAGGATTAGATACCCT3′ |
Eurofins scientific |
Custom order |
| Primer oligonucleotides for Mycoplasma screening antisense MGSO 5′TGCACCATCTGTCACTCTGTTAACCTC3′ |
Eurofins scientific |
Custom order |
| Primer oligonucleotides for generating DENV/WNV Chimera 1r: CTC TTT CCC CAG GCC TTC CAG CCA ATT TCC AAT TTT TCC GTG GTG GCG GTG AGG CGT TTA GGT CCT GCC TGC ATG ATT CCT TTG ATG |
Eurofins scientific |
Custom order |
| Primer oligonucleotides for generating DENV/WNV Chimera 2f: TGG AAG GCC TGG GGA AAG AGT ATT TTA TTT GCA CCA GAA CTC GCC AAC AAC ACC TTT GTG GTT GAT GGC CCC GAA ACA GCA GAA |
Eurofins scientific |
Custom order |
| Recombinant DNA |
| pmab vector |
Gift from Michael Diamond (Washington University in St. Louis) |
NA |
| Software and Algorithms |
| ImageJ (Image processing and analyses) |
NA |
https://www.nature.com/articles/nmeth.2089 |
| Image Studio Lite Ver 5.2 |
NA |
https://www.licor.com/bio/products/software/image_studio_lite/ |
| Graph Pad Prism 6 2D scientific graphing and biostatistics) |
NA |
https://www.graphpad.com/scientific-software/prism/ |
| Adobe Illustrator CC 2018 |
Adobe Inc. |
http://www.adobe.com |
| Microsoft PowerPoint |
Microsoft Corporation |
https://www.microsoft.com/en-us/ |
| FlowJo V10 |
FlowJo LLC |
https://www.flowjo.com/ |
| Other |
| Zeiss LSM 710 Axio Observer Zeiss Axio Observer |
ZEISS |
CRL MIC, UC Berkeley |
| Chopstick Electrode Set for EVOM2 |
World Precision Instruments |
STX3 |
| EPITHELIAL VOLTOHMMETER 2 |
World Precision Instruments |
EVOM2 |
