Antibodies |
|
IC, immunocytochemistry; WB, western blot |
Rabbit anti-G6Pase-β (IC) |
Santa Cruz Biotechnology, Dallas, USA |
Cat#sc-134714; RRID:AB_10647643
|
Rabbit anti-G6PT (WB) |
Santa Cruz Biotechnology |
Cat#sc-135479; RRID:AB_2254868
|
Donkey anti-mouse IgG-HRP (WB) |
Santa Cruz Biotechnology |
Cat#sc-2314; RRID:AB_641170
|
Donkey anti-rabbit IgG-HRP (WB) |
Santa Cruz Biotechnology |
Cat#sc-2077; RRID:AB_631745
|
Goat anti-GFAP (IC) |
AbCam, Cambridge, UK |
Cat#ab 53554; RRID:AB_880202
|
Rabbit anti-G6Pase-α (WB) |
AbCam |
Cat#ab 83690; RRID:AB_1860503
|
Rabbit anti-G6Pase-β (WB) |
AbCam |
Cat#ab 133964 |
Donkey anti-rabbit AlexaFluor 488 (IC) |
ThermoFisher, Paisley, UK |
Cat#A-21206; RRID:AB_141708
|
Donkey anti-goat AlexaFluor 633 (IC) |
ThermoFisher |
Cat#A-21082; RRID:AB_141493
|
|
Bacteria and Virus Strains |
|
Lentiviral transfer vector, FUGW |
Addgene |
Addgene#14883 |
Envelope vector, pMD2.G |
Addgene |
Addgene#12259 |
Packaging vector, pMDLg/pRRE |
Addgene |
Addgene#12251 |
Packaging vector, pRSV.Rev, |
Addgene |
Addgene#12253 |
Packaging vector, psPAX2 |
Addgene |
Addgene#12260 |
|
Chemicals, Peptides, and Recombinant Proteins |
|
ATP |
Sigma-Aldrich, Gillingham, UK |
Cat#A9187 |
2-deoxy-d-glucose |
Sigma-Aldrich |
Cat#D6134 |
AlexaFluor 568-conjugated isolectin GS-B4
|
ThermoFisher |
Cat#121412 |
Astrocyte growth medium (AGM BulletKit) |
Lonza, Slough, UK |
Cat#CC 3186 |
BAPTA |
Molekula, Dorset, UK |
Cat#20358510 |
Bovine serum albumin (BSA) |
Europa Bioproducts Ltd, Cambridge, UK |
Cat#EQBAH64 |
Cal-520AM |
AAT Stratech Scientific, Suffolk, UK |
Cat#21130 |
Dimethyl sulfoxide (DMSO) |
Sigma-Aldrich |
Cat#D2650 |
ECL Prime reagents |
GE Healthcare Life Sciences, Little Chalfont, UK |
Cat#RPN2232 |
ER Tracker Red |
ThermoFisher |
Cat#E34250 |
Fibronectin |
Merck Millipore, Watford, UK |
Cat#FC010 |
Fetal bovine serum (FBS) |
Sigma-Aldrich |
Cat#F7524, batch 094M3341 |
d-glucose-6-phosphate (G6P) |
Sigma-Aldrich |
Cat#G7879 |
Ionomycin |
Apollo Scientific, Bredbury, UK |
Cat#56092-81-0 |
Lipofectamine LTX |
ThermoFisher |
Cat#15338100 |
PAR-1 peptide, TFLLR |
Tocris, Bristol, UK |
Cat#1464 |
Probenecid |
Sigma-Aldrich |
Cat#P8761 |
Triton X-100 |
Sigma-Aldrich |
Cat#T8787 |
|
Critical Commercial Assays |
|
ATP assay kit |
AbCam |
Cat#ab113849 |
Glucose Uptake-Glo assay kit |
Promega Southampton, UK |
Cat#J1341 |
Fastlane cell cDNA kit |
QIAGEN |
Cat#215011 |
|
Experimental Models: Cell Lines |
|
293FT cells |
ThermoFisher |
Cat#R70007
|
Human astrocytes from fetal cortex |
Lonza, Slough, UK |
Cat#CC-2565 |
|
Recombinant DNA |
|
Plasmid (pEF/myc/ER FLIPglu-600uDelta13V) encoding ERglc600 |
Addgene |
Addgene#18020 |
Plasmid (pEF/myc/ER FLIPglu-30uDelta13V) encoding ERglc30 |
Addgene |
Addgene#18021 |
CCSB-Broad LentiORF-G6Pase-α |
Dharmacon, Lafayette, USA |
Cat#ccsbBroad304_04582 |
TRC human G6Pase-β shRNA |
Dharmacon |
Cat#TRCN0000051500 |
Quantitect primer assay: GAPDH |
QIAGEN, Crawley, West Sussex, UK |
Cat#Hs_GAPDH_1_SG |
Quantitect primer assay: G6Pase-β |
QIAGEN |
Cat#Hs_G6PC3_1_SG |
Quantitect primer assay: G6Pase-α |
QIAGEN |
Cat#Hs_G6PC_1_SG |
Quantitect primer assay: GPBB |
QIAGEN |
Cat#Hs_PYGB_1_SG |
Quantitect primer assay: GPMM |
QIAGEN |
Cat#Hs_PYGM_1_SG |
Quantitect primer assay: GFAP |
QIAGEN |
Cat#Hs_GFAP_1_SG |
|
Software and Algorithms |
|
Prism 5 |
GraphPad, La Jolla, USA |
https://www.graphpad.com/ |