Antibodies |
anti-ACTB |
Abcam |
Cat#ab8227 |
anti-LC3B |
Sigma-Aldrich |
Cat#L7543 |
anti-p62/SQSTM1 |
Abcam |
Cat#ab56416 |
anti-HIF1A |
BD |
Cat#610959 |
anti-HIF1B/ARNT |
BD |
Cat#611079 |
anti-HIF2A |
Novus Biologicals |
Cat#NB100-122 |
anti-BNIP3 |
Abcam |
Cat#ab109362 |
anti-TUBB |
Abcam |
Cat#ab6046 |
anti-Mouse IgG, HRP-conjugated |
MP Biomedical |
Cat#0855550 |
anti-Rabbit IgG, HRP-conjugated |
MP Biomedical |
Cat#0855676 |
anti-p62 |
MBL |
Cat#M162 |
anti-LC3 |
Novus Biological |
Cat#NB100-2220 |
anti-LC3 |
Cell Signaling Technology |
Cat#2775S |
Bacterial and virus strains |
Salmonella enterica serovar Typhimurium strain SL1344 |
Lab stock |
DSM 24522 |
Chemicals, peptides, and recombinant proteins |
Rapamycin |
Thermo-Fisher |
Cat#J62473 |
Streptomycin sulfate |
Sigma-Aldrich |
Cat#S6501 |
Phoshate buffered saline (PBS) |
Thermo-Fisher |
Cat#10010023 |
LB-Miller broth |
BD |
Cat#244610 |
Agar |
BD |
Cat#214010 |
Glycerol |
Sigma-Aldrich |
Cat#G5516-500ML |
IMDM |
Corning |
Cat#10-016-CV |
GlutaMAX |
Thermo-Fisher |
Cat#35050061 |
Penicillin/streptomycin |
Thermo-Fisher |
Cat#15140122 |
Bovine calf serum |
Cytiva |
Cat#SH30072.03 |
TRIzol reagent |
Thermo-Fisher |
Cat#15596026 |
iScript Supermix reagent |
Bio-Rad |
Cat#1708841 |
2× Power SYBR Green |
Thermo-Fisher |
Cat#4367659 |
SDS-PAGE sample buffer |
Bio-Rad |
Cat#1610747 |
HALT Protease Inhibitor |
Thermo-Fisher |
Cat#78438 |
Clarity Max ECL reagent |
Bio-Rad |
Cat#1705062 |
ProLong Gold Anti-Fade reagent |
Thermo-Fisher |
Cat#P36930
|
Critical commercial assays |
Oxodish oxygen sensor plates |
Presens |
Cat#OD24 |
EZ-10 DNAaway RNA Miniprep Kit |
Bio Basic |
Cat#BS88136 |
Dual Luciferase Assay kit |
Promega |
Cat#E1960 |
FuGENE HD |
Promega |
Cat#E2311 |
Deposited data |
HIF-1α and HIF-2α ChIP-chip profiling in intestinal epithelial cells |
NCBI GEO |
Accession#GSE43108
|
Experimental models: Cell lines |
HeLa |
Lab stock |
RRID:CVCL_0030 |
C2BBe1 |
Lab stock |
RRID:CVCL_1096 |
Colonoids |
Healthy C57BL/6 mice |
Generated in-house according to established protocols |
Experimental models: Organisms/strains |
Hif1b-flox/villin-cre |
Lab colony |
Previously generated (PMID# 24248342) |
C57BL/6J wild-type |
Lab colony |
RRID:IMSR_JAX:000664 |
ODD-luciferase |
Lab colony |
Previously described (PMID# 24412613) |
Oligonucleotides |
Primers |
This publication |
See Table S1 for sequences |
anti-HIF1A shRNA |
Functional Genomics Facility (Aurora, CO) |
Accession#TRCN0000003811 |
anti-HIF1B shRNA |
Functional Genomics Facility (Aurora, CO) |
Accession#TRCN0000003819 |
anti-HIF2A shRNA |
Functional Genomics Facility (Aurora, CO) |
Accession#TRCN0000003806 |
Recombinant DNA |
pGL4.22-VEGF-HRE::dLUC |
Chi Van Dang, via Addgene |
RRID:Addgene_128096 |
tf-LC3 |
Tamotsu Yoshimori, via Addgene |
RRID:Addgene_21074 |
Software and algorithms |
LinRegPCR |
Untergasser et al. (PMID# 34433408) |
n/a |
ClustVis |
Metsalu and Vilo (PMID# 25969447) |
n/a |