Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Recombinant DNA reagent |
pTarget Bcl11b IRES-H2Bm Cherry-neo/3pUTR |
This paper | N/A | Gene targeting vector with IRES-H2B-mCherry- loxP-neo-loxP cassette knocked into 3' UTR of Bcl11b |
| Recombinant DNA reagent |
pTarget Bcl11b dEnh-hygro |
This paper | N/A | Gene targeting vector with Enhancer replaced by hygromycin cassette |
| Recombinant DNA reagent |
FRT-PGK-gb2- hygromycin-FRT cassette |
Genebridges | Cat# A010 | |
| Recombinant DNA reagent |
MSCV IRES H2B-mCerulean |
Kueh et al., 2013 | N/A | |
| Recombinant DNA reagent |
pCL-Eco | Imgenex | Cat# NBP2-29540 | |
| Strain, strain background (mouse) |
Bcl11bYFP(neo)/mCh(neo) | This paper | N/A | Two color reporter mice generated from breeding animals homozygous for either Bcl11b YFP(neo) or Bcl11b mCh(neo). See Materials and methods for details. |
| Strain, strain background (mouse) |
Bcl11bmCh(neo)/mCh(neo) | This paper | N/A | Homozygous Bcl11b mCh(neo) reporter mice used to generate two color reporter mice. Derived from Bcl11b YFP/mCh(neo) F0 chimeric mice. See Materials and methods for details. |
| Strain, strain background (mouse) |
Bcl11bYFP/mCh(neo) | This paper | N/A | Control mice for comparing the effects of the enhancer on Bcl11b expression. Generated by targeting Bcl11b mCherry gene targeting vector to V6.5 mouse embryonic stem (ES) cells with single modified Bcl11b mCitrine dneo allele. See Materials and methods for details. |
| Strain, strain background (mouse) |
Bcl11bYFPdEnh/mCh(neo) | This paper | N/A | Two color reporter mouse with Bcl11b enhancer deleted. Generated by targeting dEnh gene target vector to V6.5 mouse ES cells with genotype Bcl11b YFP/mCh(neo). See Materials and methods for details. |
| Strain, strain background (mouse) |
Bcl11bYFPdEnh/dEnh | This paper | N/A | Homozygous deleted enhancer mice generated from Bcl11b YFP dEnh/mCh(neo) mice. See Materials and methods for details. |
| Strain, strain background (mouse) |
CD45.1 C57BL/6: B6.SJL-Ptprca Pepcb/BoyJ |
Jackson Laboratory | Stock No# 002014 | |
| Cell line (mouse) | OP9-DL1-GFP | Schmitt and Zúñiga-Pflücker, 2002 | N/A | |
| Cell line (mouse) | OP9-Mig | Schmitt and Zúñiga-Pflücker, 2002 | N/A | |
| Cell line (mouse) | OP9-DL1-hCD8 | Kueh et al., 2016 | N/A | |
| Cell line (human) | Human Phoenix-ECO | ATCC | Cat# CRL-3214 | |
| Antibody | Anti-mouse CD8a Biotin (clone 53–6.7) |
eBioscience | Cat# 13-0081-86; RRID:AB_466348 |
(1:100) |
| Antibody | Anti-mouse TCRb Biotin (clone H57-597) |
eBioscience | Cat# 13-5961-85; RRID:AB_466820 |
(1:100) |
| Antibody | Anti-mouse TCRgd Biotin (clone GL3) |
eBioscience | Cat# 13-5711-85; RRID:AB_466669 |
(1:100) |
| Antibody | Anti-mouse Ter119 Biotin (clone TER-119) |
eBioscience | Cat# 13-5921-85; RRID:AB_466798 |
(1:100) |
| Antibody | Anti-mouse NK1.1 Biotin (clone PK136) |
eBioscience | Cat# 13-5941-85; RRID:AB_466805 |
(1:100) |
| Antibody | Anti-mouse Gr-1 Biotin (clone RB6-8C5) |
eBioscience | Cat# 13-5931-86; RRID:AB_466802 |
(1:100) |
| Antibody | Anti-mouse CD11c Biotin (clone N418) |
eBioscience | Cat# 13-0114-85; RRID:AB_466364 |
(1:100) |
| Antibody | Anti-mouse CD11b Biotin (clone M1/70) |
eBioscience | Cat# 13-0112-86; RRID:AB_466361 |
(1:100) |
| Antibody | Anti-mouse CD19 Biotin (clone 1D3/6D5) |
eBioscience | Cat# 13-0193-85; RRID:AB_657658 |
(1:100) |
| Antibody | Anti-mouse CD3e Biotin (clone 145–2 C11) |
eBioscience | Cat# 13-0031-85; RRID:AB_466320 |
(1:100) |
| Antibody | Anti-human/mouse B220 Biotin (clone RA3-6B2) |
eBioscience | Cat# 13-0452-85; RRID:AB_466450 |
(1:100) |
| Antibody | Anti-mouse F4/80 Biotin (clone BM8) |
eBioscience | Cat# 13-4801-85; RRID:AB_466658 |
(1:100) |
| Antibody | Anti-mouse CD4 Biotin (clone GK1.5) |
eBioscience | Cat# 13-0041-85; RRID:AB_466326 |
(1:100) |
| Antibody | Anti-human/mouse CD44 eFluor 450 (clone IM7) |
eBioscience | Cat# 48-0441-82; RRID:AB_1272246 |
(1:300) |
| Antibody | Anti-mouse CD25 Brilliant Violet 510 (clone PC61) |
Biolegend | Cat# 102041; RRID:AB_2562269 |
(1:300) |
| Antibody | Anti-mouse CD117 (cKit) APC-eFluor 780 (clone 2B8) |
eBioscience | Cat# 47-1171-82; RRID:AB_1272177 |
(1:300) |
| Antibody | Anti-mouse HSA eFluor 450 (clone M1/69) |
eBioscience | Cat# 48-0242-82; RRID:AB_1311169 |
(1:300) |
| Antibody | Anti-mouse CD4 Brilliant Violet 510 (clone GK1.5) |
Biolegend | Cat# 100449; RRID:AB_2564587 |
(1:300) |
| Antibody | Anti-mouse CD8a APC (clone 53–6.7) |
eBioscience | Cat# 17-0081-82; RRID:AB_469335 |
(1:300) |
| Antibody | Anti-mouse TCRb APC-eFluor 780 (clone H57-597) |
eBioscience | Cat# 47-5961-82; RRID:AB_1272173 |
(1:300) |
| Antibody | Anti-mouse CD25 APC-eFluor 780 (clone PC61.5) |
eBioscience | Cat# 47-0251-82; RRID:AB_1272179 |
(1:300) |
| Antibody | Anti-mouse CD19 eFluor 450 (clone 1D3/6D5) |
eBioscience | Cat# 48-0193-82; RRID:AB_2734905 |
(1:300) |
| Antibody | Anti-mouse CD117 (cKit) APC (clone 2B8) |
eBioscience | Cat# 17-1171-82; RRID:AB_469430 |
(1:300) |
| Antibody | Anti-mouse CD45 APC-eFluor 780 (clone 30-F11) |
eBioscience | Cat# 47-0451-82; RRID:AB_1548781 |
(1:300) |
| Antibody | Anti-mouse CD25 APC (clone PC61.5) |
eBioscience | Cat# 17-0251-82; RRID:AB_469366 |
(1:300) |
| Antibody | Anti-mouse CD4 APC-eFluor 780 (clone GK1.5) |
eBioscience | Cat# 47-0041-82; RRID:AB_11218896 |
(1:300) |
| Antibody | Anti-mouse CD8a APC-eFluor 780 (clone 53–6.7) |
eBioscience | Cat# 47-0081-82; RRID:AB_1272185 |
(1:300) |
| Antibody | Anti-mouse CD45 APC (clone 30-F11) |
eBioscience | Cat# 17-0451-82; RRID:AB_469392 |
(1:300) |
| Antibody | Anti-mouse CD5 eFluor 450 (clone 53–7.3) |
eBioscience | Cat# 48-0051-82; RRID:AB_1603250 |
(1:300) |
| Antibody | Anti-mouse TCRgd APC (clone GL3) |
eBioscience | Cat# 17-5711-82; RRID:AB_842756 |
(1:300) |
| Antibody | Anti-mouse CD49b eFluor 450 (clone DX5) |
eBioscience | Cat# 48-5971-82; RRID:AB_10671541 |
(1:300) |
| Antibody | Anti-mouse NK1.1 APC (clone PK136) |
eBioscience | Cat# 17-5941-82; RRID:AB_469479 |
(1:300) |
| Antibody | Anti-mouse CD3e APC-eFluor 780 (clone 145–2 C11) |
eBioscience | Cat# 47-0031-82; RRID:AB_11149861 |
(1:300) |
| Antibody | Anti-mouse TCRb eFluor 450 (clone H57-597) |
eBioscience | Cat# 48-5961-82; RRID:AB_11039532 |
(1:300) |
| Antibody | Anti-mouse CD49b Biotin (clone DX5) |
eBioscience | Cat# 13-5971-82; RRID:AB_466825 |
(1:300) |
| Antibody | Anti-mouse CD62L APC (clone MEL-14) |
eBioscience | Cat# 17-0621-82; RRID:AB_469410 |
(1:300) |
| Antibody | Anti-mouse CD45.2 Brilliant Violet 510 (clone 104) |
Biolegend | Cat# 109837; RRID:AB_2561393 |
(1:300) |
| Antibody | Anti-mouse CD4 eFluor 450 (clone GK1.5) |
eBioscience | Cat# 48-0041-82; RRID:AB_10718983 |
(1:300) |
| Antibody | Anti-mouse CD45 eFluor 450 (clone 30-F11) |
eBioscience | Cat# 48-0451-82; RRID:AB_1518806 |
(1:300) |
| Antibody | Streptavidin PerCP-Cyanine5.5 |
Biolegend | Cat# 405214; RRID:AB_2716577 |
(1:300) |
| Antibody | Streptavidin Brilliant Violet 510 |
Biolegend | Cat# 405234 | (1:300) |
| Peptide, recombinant protein |
Recombinant Human Flt3-Ligand |
PeproTech | Cat# 300–19 | |
| Peptide, recombinant protein |
Recombinant Human IL-7 | PeproTech | Cat# 200–07 | |
| Peptide, recombinant protein |
Recombinant Human Stem Cell Factor (SCF) |
PeproTech | Cat# 300–07 | |
| Peptide, recombinant protein |
Recombinant Mouse IL-6 | eBioscience | Cat# 14-8061-62 | |
| Peptide, recombinant protein |
Recombinant Mouse Stem Cell Factor (SCF) |
eBioscience | Cat# 34-8341-82 | |
| Peptide, recombinant protein |
Recombinant Mouse IL-3 | eBioscience | Cat# 14-8031-62 | |
| Peptide, recombinant protein |
Retronectin | Takara | Cat# T100B | |
| Peptide, recombinant protein |
DL1-extIgG Protein | Varnum-Finney et al., 2000 | N/A | |
| Software, algorithm |
FlowJo (v10.0.8) | Tree Star | N/A | |
| Software, algorithm |
MATLAB (R2016a) | MathWorks | N/A | |
| Other | FuGENE 6 Transfection Reagent |
Promega | Cat# E2691 | |
| Other | MACS Streptavidin Microbeads |
Miltenyi Biotec | Cat# 130-048-101 | |
| Other | LS Columns | Miltenyi Biotec | Cat# 130-042-401 | |
| Other | 250mm-diameter PDMS circular micromesh arrays |
Microsurfaces Pty Ltd | Cat# MMA-0250-100-08-01 |