Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Mus musculus) | eLrrc8a KO (Lrrc8afl/fl) | This paper | Sah lab | SWELL1 is a regulator of adipocyte size, insulin signaling, and glucose homeostasis (Zhang et al., 2017) |
| Strain, strain background (Mus musculus) | CDH5-Cre | Gift from Dr. Kaikobad Irani | Vikram, A, Nature Communications 2016 | |
| Strain, strain background (Mus musculus) | Rosa26-tdTomato | Jackson lab | Jax 007914 RRID:IMSR_JAX:007914 |
|
| Other | HUVECs | ATCC | Cat#CRL-2922 RRID:CVCL_3901 |
Human primary cells |
| Biological sample (Mus musculus) | Endothelial primary cell | Lrrc8afl/fl | Freshly isolated from Lrrc8afl/fl mice | |
| Antibody | Anti-ß-actin (rabbit monoclonal) | Cell Signaling | Cat#8457s, RRID:AB_10950489 |
WB (1:2000) |
| Antibody | Anti-total Akt (rabbit monoclonal) |
Cell Signaling | Cat#4685s, RRID:AB_2225340 | WB (1:1000) |
| Antibody | Anti-Akt1 (rabbit monoclonal) |
Cell Signaling | Cat#2938s, RRID:AB_915788 | WB (1:1000) |
| Antibody | Anti-p-eNOS (rabbit monoclonal) |
Cell Signaling | Cat#9571, RRID:AB_329837 | WB (1:1000) IF (1:200) |
| Antibody | Anti-p-eNOS (rabbit polyclonal) |
Invitrogen | Cat#PA5-104858, RRID: AB_2816331 | IF (1:200) |
| Antibody | Anti-Akt2 (rabbit monoclonal) |
Cell Signaling | Cat#3063s, RRID:AB_2225186 | WB (1:1000) |
| Antibody | Anti-p-AS160 (rabbit polyclonal) |
Cell Signaling | Cat#4288s, RRID:AB_10545274 | WB (1:1000) |
| Antibody | Anti-total eNOS (rabbit polyclonal) |
Cell Signaling | Cat#32027, RRID:AB_2728756 | WB (1:1000) |
| Antibody | Anti-PECAM (rabbit monoclonal) |
Sigma | Cat#SAB4502167 RRID:AB_10762600 |
WB (1:1000) |
| Antibody | Anti-VEGFR2 (goat polyclonal) |
R and D Systems | Cat#BAF357 RRID:AB_356414 |
WB (1:1000) |
| Antibody | Anti- pS6 ribosomal (rabbit monoclonal) |
Cell Signaling | Cat#5364s, RRID:AB_10694233 | WB (1:2000) |
| Antibody | Anti- GAPDH (rabbit monoclonal) |
Cell Signaling | Cat#5174s, RRID:AB_1062202 |
WB (1:2000) |
| Antibody | Anti-pErk1/2 (rabbit polyclonal) |
Cell Signaling | Cat#9101s, RRID:AB_331772 |
WB (1:1000) |
| Antibody | Anti-Total Erk1/2 (rabbit polyclonal) |
Cell Signaling | Cat#9102s, RRID:AB_330744 |
WB (1:1000) |
| Antibody | Anti-Grb2 (mouse monoclonal) |
BD | Cat#610111s, RRID:AB_397517 |
WB (1:1000) |
| Antibody | Anti-Grb2 (rabbit monoclonal) |
Santa Cruz | Cat#sc-255, RRID:AB_631602 |
WB (1:1000) |
| Antibody | Anti-LRRC8A (rabbit polyclonal) |
Pacific antibodies | Custom made | Epitope: QRTKSRIEQGIVDRSE, WB (1:1000), LRRC8A is a glucose sensor regulating ß-cell excitability and systemic glycemia (Kang et al., 2018) |
| Antibody | Anti-CD31 (rabbit polyclonal) |
Thermo Fisher | Cat#MA3105, RRID:AB_223592 |
IF (1:100) |
| Antibody | Anti-IgG (normal mouse IgG) |
Santa Cruz | Cat#sc-2027, RRID:AB_737197 |
WB (1:1000) |
| Antibody | Anti-rabbit-HRP | BioRad | Cat#170–6515, RRID:AB_11125142 |
WB (1:10,000) |
| Antibody | Anti-mouse-HRP | BioRad | Cat#170–5047, RRID:AB_11125753 |
WB (1:10,000) |
| Transfected construct (human) | siLrrc8a | Invitrogen | Cat#4392420 | Assay ID: s32107 Transfected construct human |
| Transfected construct (human) | Non-targeting control | Invitrogen | Cat#4390846 | Transfected construct human |
| Transfected construct (human) | siPORTamine | Invitrogen | Cat#AM4503 | |
| Transfected construct (human) | Opti-MEM | Invitrogen | Cat#11058–021 | |
| Recombinant DNA reagent | Ad5-U6-hLrrc8a-shRNA-mCherry | Vector biolabs | shADV- 653 214592 |
|
| Recombinant DNA reagent | Ad5-U6-Scr-shRNA-mCherry | Vector biolabs | 3086 | |
| Commercial assay or kit | RNA isolation (PureLink RNA mini kit) | Invitrogen | 12183018A | |
| Software, algorithm | GraphPad Prism8 | La Jolla California USA, http://www.graphpad.com’ RRID:SCR_002798 | ||
| Software, algorithm | Fiji, ImageJ |
Schindelin et al., 2012
(PMID:22743772) |
RRID:SCR_002285 | |
| Other | DAPI stain | Thermofisher | Cat#T3605 | 1:10,000 |