| Antibodies |
| APC Anti-mouse CD3ε (17A2) |
BioLegend |
Cat #100236 |
| FITC Anti-mouse CD3ε (17A2) |
BioLegend |
Cat #100204 |
| PE Anti-mouse CD3ε (17A2) |
BioLegend |
Cat #100206 |
| APC Anti-mouse CD4 (GK1.5) |
BioLegend |
Cat #100412 |
| FITC Anti-mouse CD4 (GK1.5) |
BioLegend |
Cat #100406 |
| PE Anti-mouse CD4 (GK1.5) |
BioLegend |
Cat #100408 |
| APC Anti-mouse CD5 (53–7.3) |
BioLegend |
Cat #100626 |
| FITC Anti-mouse CD5 (53–7.3) |
BioLegend |
Cat #100606 |
| PE Anti-mouse CD5 (53–7.3) |
BioLegend |
Cat #100608 |
| APC Anti-mouse CD8α (53–6.7) |
eBioscience |
Cat #17–0081–82 |
| FITC Anti-mouse CD8α (53–6.7) |
eBioscience |
Cat #11–0081–85 |
| PE Anti-mouse CD8α (53–6.7) |
eBioscience |
Cat #12–0081–83 |
| APC Anti-mouse CD11b (M1/70) |
eBioscience |
Cat #17–0112–82 |
| APC efluor780 Anti-mouse CD11b (M1/70) |
eBioscience |
Cat #47–0112–82 |
| FITC Anti-mouse CD11b (M1/70) |
eBioscience |
Cat #12–0112–83 |
| PE Anti-mouse CD11b (M1/70) |
eBioscience |
Cat #11–0112–85 |
| PE/Cy7 Anti-mouse CD16/32 (FcγRII/III; 93) |
BioLegend |
Cat #101318 |
| PerCP/Cy5.5 Anti-mouse CD16/32 (FcγRII/III; 93) |
BioLegend |
Cat #101324 |
| Biotin Anti-mouse CD34 (RAM34) |
eBioscience |
Cat #13–0341–85\ |
| eFluor660 Anti-mouse CD34 (RAM34) |
eBioscience |
Cat #50–0341–82 |
| Alexa Fluor 700 Anti-mouse CD34 (RAM34) |
eBioscience |
Cat #56–0341–82 |
| FITC Anti-mouse CD34 (RAM34) |
eBioscience |
Cat #11–0341–85 |
| FITC Anti-mouse CD43 (R2/60) |
eBioscience |
Cat #11–0431–85 |
| PE Anti-mouse CD43 (R2/60) |
eBioscience |
Cat #12–0431–83 |
| APC eFluor 780 Anti-mouse CD45.1 (A20) |
eBioscience |
Cat #47–0453–82 |
| Alexa Fluor 700 Anti-mouse CD45.2 (104) |
BioLegend |
Cat #109822 |
| FITC Anti-mouse CD45.2 (104) |
BioLegend |
Cat #109806 |
| APC Anti-mouse CD45R (B220) (RA3–6B2) |
eBioscience |
Cat #17–0452–83 |
| FITC Anti-mouse CD45R (B220) (RA3–6B2) |
eBioscience |
Cat #11–0452–85 |
| PE Anti-mouse CD45R (B220) (RA3–6B2) |
eBioscience |
Cat #12–0452–85 |
| PerCP-Cyanine5 Anti-mouse CD45R (B220) (RA3–6B2) |
eBioscience |
Cat #45–0452–82 |
| APC Anti-mouse CD48 (HM48–1) |
BioLegend |
Cat #103412 |
| FITC Anti-mouse CD48 (HM48–1) |
BioLegend |
Cat #103404 |
| PE Anti-mouse CD48 (HM48–1) |
BioLegend |
Cat #103406 |
| PE/Cy7 Anti-mouse CD48 (HM48–1) |
BioLegend |
Cat #103424 |
| FITC Anti-mouse CD71 (R17217) |
eBioscience |
Cat #11–0711–85 |
| APC Anti-mouse CD117 (cKit) (2B8) |
eBioscience |
Cat #17–1171–83 |
| APC eFluor 780 Anti-mouse CD117 (cKit) (2B8) |
eBioscience |
Cat #47–1711–82 |
| PE-Cyanine7 Anti-mouse CD117 (cKit) (2B8) |
eBioscience |
Cat #25–1711–82 |
| PE Anti-mouse CD127 (IL7Rα; A7R34) |
BioLegend |
Cat #135010 |
| APC Anti-mouse CD150 (TC15–12F12.2) |
BioLegend |
Cat #115910 |
| PE Anti-mouse CD150 (TC15–12F12.2) |
BioLegend |
Cat #115904 |
| PE-Cyanine7 Anti-mouse CD150 (TC15–12F12.2) |
BioLegend |
Cat #115914 |
| APC Anti-mouse Ter119 (TER-119) |
Biolegend |
Cat #116212 |
| FITC Anti-mouse Ter119 (TER-119) |
Biolegend |
Cat #116206 |
| PE Ter119 (TER-119) |
Biolegend |
Cat #116208 |
| APC Anti-mouse Sca-1 (D7, E13–161.7)\ |
eBioscience |
Cat #17–5981–82 |
| Alexa Fluor 700 Anti-mouse Sca-1 (D7, E13–161.7) |
eBioscience |
Cat #56–5981–82 |
| FITC Anti-mouse Sca-1 (D7, E13–161.7) |
eBioscience |
Cat #11–5981–85 |
| PerCp-Cyanine5.5 Anti-mouse Sca-1 (D7, E13–161.7) |
eBioscience |
Cat #45–5981–82 |
| APC Anti-mouse Gr-1 (RB6–8C5) |
BioLegend |
Cat #108412 |
| FITC Anti-mouse Gr-1 (RB6–8C5) |
BioLegend |
Cat #108406 |
| PE Anti-mouse Gr-1 (RB6–8C5) |
BioLegend |
Cat #108408 |
| PE/Cy7 Anti-mouse Gr-1 (RB6–8C5) |
BioLegend |
Cat #108416 |
| APC Anti-mouse IgM (II/41) |
eBioscience |
Cat #17–5790–82 |
| PE Anti-mouse IgM (II/41) |
eBioscience |
Cat #12–5790–83 |
| Streptavidin-PECy7 |
BioLegend |
Cat #405206 |
| Anti-Ubiquitin (P4D1) |
Cell Signaling |
Cat #3933S |
| Anti-K48-Polyubiquitin (polyclonal) |
Cell Signaling |
Cat# 4289S |
| Anti-c-Myc (polyclonal) |
Cell Signaling |
Cat# 9402S |
| Anti-phos-Eif2α (S51) |
Cell Signaling |
Cat #9721S |
| Anti-β-Actin (AC74) |
Sigma |
Cat #A2228 |
| Anti-Gapdh (14C10) |
Cell Signaling |
Cat #2118S |
| HRP-linked anti-rabbit IgG |
Cell Signaling |
Cat #7074S |
| HRP-linked anti-mouse IgG |
Cell Signaling |
Cat #7076S |
| Chemicals, Peptides, and Recombinant Proteins |
| PIPC |
GE Healthcare Biosciences Corp |
Cat #27473201 |
| Anti-mouse CD117 microbeads |
Miltenyi |
Cat #130–091–224 |
| TCA |
Sigma |
Cat #T6399 |
| Triton X-100 |
CalBiochem |
Cat #648466 |
| LDS loading buffer |
Life Technologies |
Cat #NP007 |
| SDS |
Life Technologies |
Cat #NP0060 |
| Glycine |
Sigma |
Cat #50046 |
| Tetraphenylethene maleimide (TMI) |
Dr. Yuning Hong |
Custom Synthesized |
| OP-Puro |
Medchem Source |
Custom Synthesized |
| SYPRO Ruby Protein Gel Stain |
Bio-Rad |
Cat #1703125 |
| 2-mercaptoethanol |
Sigma |
Cat #M3148 |
| SYBR green |
Roche |
Cat #4707516001 |
| Critical Commercial Assays |
| SuperSignal West Femto or Pico PLUS chemiluminescence kit |
Thermo Scientific |
Cat #34095 |
| Click-iT Cell Reaction Buffer Kit |
Life Technologies |
Cat #C10269
|
| Alexa Fluor 555-conjugated azide |
Life Technologies |
Cat #A20012 |
| Proteasome-Glo chymotrypsin-like cell based assay |
Promega |
Cat #G8660 |
| RNeasy Plus Micro Kit |
QIAGEN |
Cat #74034 |
| RT2 First Strand Kit |
QIAGEN |
Cat #330404 |
| Deposited Data |
| RNA-sequencing data (Aarssti/sti and Aars+/+ HSCs) |
GEO |
GSE141008 |
| Experimental Models: Organisms/Strains |
| Mouse: B6: C57BL/6J |
The Jackson Laboratory |
Cat #000664 |
| Mouse: B6SJL: B6.SJL |
The Jackson Laboratory |
Cat #002014 |
| Mouse: Ptenfl/fl
|
The Jackson Laboratory |
Cat #006440 |
| Mouse: Mx1-Cre |
The Jackson Laboratory |
Cat #003556 |
| Mouse: Rpl24Bst/+
|
The Jackson Laboratory |
Cat #000516 |
| Mouse: Aarssti/sti
|
The Jackson Laboratory |
Cat #002560 |
| Mouse: UbG76V-GFP
|
The Jackson Laboratory |
Cat #008111 |
| Mouse: Mycfl
|
The Jackson Laboratory |
Cat #011123 |
| Oligonucleotides |
|
Myc forward |
IDT |
5′- CTTCTCTCCTTCCTCGGACTC-3′ |
|
Myc reverse |
IDT |
5′- GGAGATGAGCCCGACTCCGACCTC-3′ |
| β-Actin forward |
IDT |
5′-CGTCGACAACGGCTCCGGCATG-3′ |
| β-Actin reverse |
IDT |
5′-GGGCCTCGTCACCCACATAGGAG-3′ |
| Software and Algorithms |
| FlowJo |
FlowJo, LLC |
N/A |
| FACSDiva |
BD Bioscience |
N/A |
| Prism 8 |
GraphPad |
N/A |
| ImageJ |
NIH |
N/A |
| Image Lab 6.0.1 |
Bio-Rad |
N/A |
| TopHat 1.4.1 |
JHU CCB |
N/A |
| PRINSEQ Lite 0.20.3 |
N/A |
N/A |
| Htseq-count 0.7.1 |
N/A |
N/A |
| Bioconductor package DESeq2 1.6.3 |
Bioconductor |
N/A |
