Antibodies |
Rat anti-C-peptide |
Developmental Studies Hybridoma Bank at the University of Iowa |
GN-ID4; RRID:AB_2255626 |
Mouse anti-NKX6.1 |
Developmental Studies Hybridoma Bank at the University of Iowa |
F55A12; RRID:AB_532379 |
Rabbit anti-Beta Tubulin |
Cell Signaling Technology |
2128; RRID:AB_823664 |
Mouse anti-PGK1/2 |
Santa Cruz Biotechnology |
SC-48342; RRID:AB_628116 |
Rabbit anti-GAPDH |
Cell Signaling Technology |
5174; RRID:AB_10622025 |
Rabbit anti-PC |
Sigma Aldrich |
HPA043922; RRID:AB_2678732 |
Rabbit anti-PCK2 |
Cell Signaling Technology |
8565; RRID:AB_11217628 |
Biological Samples |
Human Cadaveric Islets |
See Table S1
|
N/A |
Chemicals, Peptides, and Recombinant Proteins |
Phenylacetate |
Sigma Aldrich |
P16621 |
Tris-HCl pH 8.5 |
VWR |
100219-630 |
Disposable spinner flasks |
Corning |
CLS3578 |
mTeSR basal medium |
StemCell Technologies, Inc. |
85850 |
MCDB131 Basal Medium |
Sigma Aldrich |
M8537 |
Fetal Bovine Serum |
Life Technologies |
16000044 |
Fluo-4 |
Life Technologies |
F14217 |
Oligomycin A |
Fisher Scientific |
49545510MG |
Disuccinyl Suberate |
Life Technologies |
4449754 |
U-13C Glucose |
Cambridge Isotope Laboratories |
CLM-1396-0.25 |
L-Glutamine |
Sigma Aldrich |
G3126 |
TSQ |
Fisher Scientific |
50850395 |
Propidium Iodide |
Sigma Aldrich |
P4864 |
Sodium Fluoride |
Sigma Aldrich |
919 |
96 well V-bottom plates |
Fisher Scientific |
12565460 |
methyl Succinate |
Sigma Aldrich |
P4241-50EA |
Methyl-pyruvate |
VWR |
AAA13966 |
Glyceraldehyde |
Sigma Aldrich |
49800 |
Glyceric Acid |
Santa Cruz Biotechnology |
SC-234508 |
Dihydroxyacetone |
Santa Cruz Biotechnology |
SC499029 |
B-hydroxybutyrate |
Sigma Aldrich |
166898 |
Ketoisocaproate |
VWR |
AAH60076-03 |
D-3-Phosphoglyceric Acid |
Santa Cruz Biotechnology |
SC-214793B |
D-2-Phosphoglyceric Acid |
Sigma Aldrich |
79470 |
Tolbutamide |
Sigma Aldrich |
T0891 |
Diazoxide |
Sigma Aldrich |
D9035 |
Forskolin |
Fisher Scientific |
NC9084630 |
MitoTracker Green FM |
Life Technologies |
M7514 |
RIPA Lysis Buffer |
Fisher Scientific |
P189901 |
Glucose |
Sigma Aldrich |
G7021 |
Potassium Chloride |
Sigma Aldrich |
P9541 |
Sodium Chloride |
Sigma Aldrich |
S3014 |
Calcium Chloride |
Sigma Aldrich |
C7902 |
Magnesium Sulfate |
Sigma Aldrich |
M7506 |
Sodium Phosphate, Dibasic |
Sigma Aldrich |
S3264 |
Potassium Phosphate, Monobasic |
Sigma Aldrich |
P9791 |
Sodium Bicarbonate |
Sigma Aldrich |
S5761 |
HEPES Buffer |
Life Technologies |
15630080 |
Dynabeads Protein G |
Life Technologies |
10003D |
Bovine Serume Albumin, Fatty acid-free, Fraction V |
EMD Millipore |
126575 |
2-NBDG |
Life Technologies |
N13195 |
Critical Commercial Assays |
Seahorse 24 Xf Mito Fuel Stress Test |
Agilent |
103015-100 |
Seahorse 24 well islet capture plate |
Agilent |
101122-100 |
PGK1 Activity Assay |
Biovision |
K194 |
GAPDH enzyme activity assay |
Abcam |
ab204732 |
PEP colorimetric Assay |
Sigma Aldrich |
MAK102-1KT |
Alpco Human Insulin ELISA |
APCO Diagnostics |
80-INSHUU-E10.1 |
Experimental Models: Cell Lines |
HUES8 hES |
Derived In-house |
N/A |
1016 iPS |
Columbia University |
N/A |
Oligonucleotides |
B2M Forward: GCTGGGTAGCTCTAAACAATGTATTCA |
This Paper adapted from Phillips et al., 2014
|
N/A |
B2M Reverse: CCATGTACTAACAAATGTCTAAAATGGT |
This Paper adapted from Phillips et al., 2014
|
N/A |
B2M FAM Probe: CAGCAGCCTATTCTGC |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMajArc Forward: CTGTTCCCCAACCTTTTCCT |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMajArc Reverse: CCATGATTGTGAGGGGTAGG |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMajArc FAM Probe: GACCCCCTAACAACCCCC |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMinArc Forward: CTAAATAGCCCACACGTTCCC |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMinArc Reverse: AGAGCTCCCGTGAGTGGTTA |
This Paper adapted from Phillips et al., 2014
|
N/A |
mtMinArc FAM Probe: CATCACGATGGATCACAGGT |
This Paper adapted from Phillips et al., 2014
|
N/A |
Recombinant DNA |
Empty Myc-DDK Lentivirus Plasmid |
Origene |
PS100064;http://www.origene.com/catalog/vectors/lentiviral-gene-expression-vectors/ps100064/plenti-c-myc-ddk-lentiviral-gene-expression-vector
|
GAPDH Myc-DDK Lentivirus Plasmid |
Origene |
RC202309L1; https://www.origene.com/cDNA_clone/LentiORF/NM_002046/RC202309L1V/GAPDH.aspx
|
PGK1 Myc-DDK Lentivirus Plasmid |
Origene |
RC211172L1 |
Software and Algorithms |
Prism |
Graphpad |
N/A |
Wave |
Agilent |
N/A |
Zen |
Zeiss |
N/A |
ImageJ |
National Institutes of Health |
https://imagej.nih.gov/ij/ |
Flowjo |
Becton, Dickinson and Company |
N/A |
Other |
AxioZoom 16 |
|
N/A |
BioRep Dynamic Perifusion System |
|
N/A |
Seahorse XFe 24 well Analyzer |
|
N/A |