Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) |
C57Bl6/J | Jax | Cat# JAX:000664, RRID:IMSR_JAX:000664 | Used for cell line transplantation |
| Genetic Reagent (M. musculus) |
LSL-KrasG12D/+
Trp53flox/flox Pdx-1-Cre LSL-tdTomato; KP-/-CT |
Bardeesy et al., 2006
PMID:16585505 |
Mice from a mixed 129/Sv and C57Bl6/J genetic background as well as pure C57Bl6/J mice were used. Both sexes of mice were included in experiments. | |
| Genetic Reagent (M. musculus) |
LSL-KrasG12D/+
Trp53R172H/+ Pdx-1-Cre LSL-tdTomato; KPCT |
Hingorani et al., 2005
PMID:15894267 |
Mice from a mixed 129/Sv and C57Bl6/J genetic background as well as pure C57Bl6/J mice were used. Both sexes of mice were included in experiments. | |
| Genetic Reagent (M. musculus) |
β-actin-GFP | Jax | Cat# JAX:006567, RRID:IMSR_JAX:006567 | PSCs were isolated from β-actin-GFP mice in a C57Bl6/J background. |
| Cell line (M. musculus) |
PSC |
Danai et al., 2018
PMID:29925948 |
Isolated from β-actin-GFP mice in a C57Bl6/J background (Jax 006567). PSCs were immortalized with TERT and SV40 largeT after several passages. | |
| Cell line (M. musculus) |
AL1376 |
Sullivan et al., 2018
PMID:29941931 |
Isolated from KP-/-CT mouse PDAC tumor in a C57Bl6/J background. | |
| Cell line (M. musculus) |
Organoid | This paper | Isolated from KP-/-CT mouse PDAC tumor in a C57Bl6/J background | |
| Biological sample (Homo sapiens) |
Human pancreatic cancer tissue microarray | Biomax | PA961e | |
| Antibody | α-SMA (mouse monoclonal) | Sigma | Cat# F3777, RRID:AB_476977 | IF (1:500) |
| Antibody | CK19 (Rabbit monoclonal) |
Abcam | Cat# ab133496, RRID:AB_ 11155282 |
IF (1:100) |
| Antibody | tdTomato (Rabbit polyclonal) | Rockland | Cat# 600-401-379, RRID:AB_2209751 | IF (1:500) |
| Antibody | Pyruvate Carboxylase (Rabbit polyclonal) |
Santa Cruz | Cat# sc-67021, RRID:AB_2283532 | IHC (1:50) |
| Antibody | Pyruvate Carboxylase (Mouse monoclonal) |
Santa Cruz | Cat# sc-271493, RRID:AB_10649369 | WB (1:100) |
| Antibody | Malic Enzyme 1 (Rabbit polyclonal) | Proteintech | Cat# 16619-1-AP, RRID:AB_2143821 | IHC (1:200) WB (1:250) |
| Antibody | β-actin (Rabbit monoclonal) | Cell Signaling Technologies | Cat# 8457, RRID:AB_10950489 | WB (1:10,000) |
| Antibody | CD16/CD32 (Rat monoclonal) | Thermo Fisher | Cat# 14-0161-82, RRID:AB_467133 | FACS (10uL) |
| Antibody | CD45-APC-Cy7 (Rat monoclonal) |
BD | Cat# 557659, RRID:AB_396774 | FACS (1:100) |
| Antibody | Puromycin (Mouse monoclonal) |
Sigma | MABE343 | WB (1:25,000) |
| Antibody | Vinculin (Mouse monoclonal) |
Abcam | Cat# ab18058, RRID:AB_444215 | WB (1:1000) |
| Commercial Assay or Kit | Protein A Antibody Purification Kit | Sigma | PURE1A | |
| Commercial Assay or Kit | RNAqueous-Micro Total RNA Isolation Kit | Life Technologies | AM1931 | |
| Commercial Assay or Kit | iScript cDNA Synthesis Kit | Bio-Rad Laboratories | 1708890 | |
| Commercial Assay or Kit | Pierce BCA Protein Assay Kit | Pierce | 23225 | |
| Commercial Assay or Kit | Amaxa Basic Nucleofector Kit for Primary Mammalian Epithelial Cells | Amaxa | VPI-1005 | |
| Recombinant DNA Reagent | pUSPmNG (plasmid) |
Li et al., 2019
PMID:31694929 |
U6 sgRNA PGK with mNeonGreen to express sgRNAs for PC and ME1 double knockout cell lines | |
| Recombinant DNA Reagent | LentiCRISPRv2 (plasmid) |
Sanjana et al., 2014
PMID:25075903 |
Lentiviral construct to create PC and ME1 knockout cells | |
| Recombinant DNA Reagent | Modified LentiCRISPRv2 (plasmid) |
Horlbeck et al., 2016
PMID:27661255 |
Modified dCas9-KRAB fusion protein to create PC and ME1 knockdown cells | |
| Recombinant DNA Reagent | pLV-Hygro-EFS (plasmid) |
Vectorbuilder | Custom lentiviral construct for re-expressing PC or ME1 cDNA under a CMV promoter | |
| Peptide, recombinant protein | EGF | Thermo Fisher | PMG8041 | |
| Peptide, recombinant protein | FGF | Peprotech | 100-26 | |
| Peptide, recombinant protein | Gastrin I | TOCRIS | 3006 | |
| Peptide, recombinant protein | Noggin | Peprotech | 250-38 | |
| Chemical compound, drug | Collagenase I | Worthington Biochemical | LS004194 | |
| Chemical compound, drug | Collagenase XI | Sigma | C9407 | |
| Chemical compound, drug | Collagenase P | Sigma | 11213865001 | |
| Chemical compound, drug | Dispase II | Roche | 04942078001 | |
| Chemical compound, drug | TrypLE Express | Thermo Fisher | 12605-010 | |
| Chemical compound, drug | DNAse I Type II |
Sigma | D4527 | |
| Chemical compound, drug | DNAse I Type IV |
Sigma | D5025 | |
| Chemical compound, drug | GlutaMAX | Thermo Fisher | 35050 | |
| Chemical compound, drug | HEPES | Thermo Fisher | 15630 | |
| Chemical compound, drug | TGF-b inhibitor A-83-01 |
TOCRIS | 2939 | |
| Chemical compound, drug | Rho Kinase Inhibitor Y-27632 | Sigma | Y0503 | |
| Chemical compound, drug | N-Acetylcysteine (NAC) | Sigma | A9165 | |
| Chemical compound, drug | Nicotinamide | Sigma | N0636 | |
| Chemical compound, drug | B-27 supplement | Thermo Fisher | 17504 | |
| Chemical compound, drug | Methoxamine (MOX) reagent | ThermoFisher | TS-45950 | |
| Chemical compound, drug | N–methyl–N–(tert–butyldimethylsilyl)trifluoroacetamide + 1% tert–Butyldimethylchlorosilane | Sigma | 375934 | |
| Chemical compound, drug | Pyridine | Sigma | 270407 | |
| Chemical compound, drug | Nycodenz | VWR | 100356-726 | |
| Chemical compound, drug | Sulforhodamine B | Sigma | 230162 | |
| Chemical compound, drug | SYBR Green Master Mix | Sigma | L6544 | |
| Software, algorithm |
Prism | GraphPad | Statistical analysis and graphing | |
| Software, algorithm | FlowJo | BD | Flow cytometry data analysis | |
| Other | SYTOX Red Dead Cell Stain | Life Technologies | S34859 | FACS (1:1000) |
| Other | Advanced DMEM/F12 | Thermo Fisher | 12634 | |
| Other | DMEM without pyruvate | Corning | 10-017-CV | |
| Other | GBSS | Sigma | G9779 | |
| Other | Growth factor reduced (GFR) matrigel | Corning | 356231 | |
| Other | Plastic coverslips | Thermo | 174985 | |
| Other | Flow cytometry staining buffer | Thermo Fisher | 00-4222-57 |