Journal: bioRxiv

Article Title: Tbet promotes NK cell egress from the bone marrow and CXCR6 expression in immature NK cells

doi: 10.1101/583575

Figure Lengend Snippet: Leukocytes were isolated from Tbx21 +/+ (WT; blue) and Tbx21 -/- (red) bone marrow and iNK, mNK1 and mNK2 cell subsets were screened by flow cytometry for differentially expressed proteins. Proteins examined were selected based on differential expression of their transcripts between Tbx21 +/+ and Tbx21 -/- in the RNASeq screen. Representative histograms and summary graphs for (a) CCR2, (b) CD49a, (c) CD62L, (d) CD69, (e) CD127, (f) cKit, (g) CXCR6, (h) CX3CR1, and (i) S1PR5 are shown. n = 6 mice per group, means and SD are shown. Significance was determined using two-sample, one-tailed t tests with Holm-Sidak correction. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The following anti-mouse antibodies were used: anti-CCR2-PE-Cyanine7 (clone SA203G11, Biolegend, London, UK), anti-CD3-FITC (17A2, Biolegend), anti-CD3-biotin (clone 145-2C11, Sony Biotechnology, Surrey, UK), anti-CD4-BV786 (clone RM4-5, BD Horizon), anti-CD8α-FITC (clone 53-6.7, Biolegend), anti-CD8α-biotin (clone 53-6, Sony), anti-CD11b-APC (clone M1/70, Biolegend), anti-CD11b-PE (clone M1/70, Biolegend), anti-CD11b-AlexaFluor700 (clone M1/70, BD Pharmingen), anti-CD19-FITC (clone 6D5, Biolegend), anti-CD19-PerCP-Cyanine5.5 (clone 6D5, Biolegend), anti-CD19-biotin (clone 6D5, Sony), anti-CD27-APC (clone LG.3A10, Biolegend), anti-CD27-APCeFluor780 (clone LG.7F9, eBioscience, San Diego, CA), anti-CD27-PE-Dazzle594 (clone LG.3A10, Biolegend), anti-CD27-APC-Cyanine7 (clone LG.7F9, eBio-science), anti-CD45-BV510 (clone 30-F11, Biolegend), anti-CD45-APC-Cyanine7 (clone 30-F11, BD Pharmingen), anti-CD45.2 PE-eFluor610 (clone 104, eBioscience), anti-CD45.1-PE-Cyanine7 (clone A20, Biolegend), anti-CD49a-BUV395 (clone Ha31/8, BD Bioscience), anti-CD49b-BV510 (clone HMα2, BD Optibuild), anti-CD62L-PerCP-Cyanine5.5 (clone MEL-14, Biolegend), anti-CD69-PerCP-Cyanine5.5 (clone H1.2F3, Biolegend), anti-CD117(cKit)-BV510 (clone 2B8, Biolegend), anti-CD117 (ckit)-APC-Cyanine7 (clone ACK2, Sony), anti-CD122 (IL-2Rβ)-eFluor450 (clone TM-β1, eBioscience), anti-CD122 (IL-2R-β)-APC (clone TM-β1, Biolegend), anti-CD127 (IL-7Rα)-PE (clone A7R34, eBioscience), anti-CD127 (IL-7Rα)-PE-Cyanine7 (clone A7R34, Sony), anti-CD135 (Flt3)-PerCP-eFluor710 (clone A2F10, eBioscience), anti-CD135 (Flt3)-PE (clone A2F10, Sony), anti-CD244.2-PE-Cyanine7 (clone m2B4 (B6)458.1, Biolegend), anti-CX3CR1-BV510 (clone SA011F11, Biolegend), anti-CXCR6-PE-Cyanine7 (clone SA051D1, Biolegend), anti-Gr-1-FITC (clone RB6-8C5, Biolegend), anti-Gr-1-APC (clone RB6-8C5, Biolegend), anti-GR1-biotin (clone RB6-8C5, BD Pharmingen), anti-integrin α4β7-BV421 (clone DATK32, BD Horizon), anti-NK1.1-FITC (clone PK136, Biolegend), anti-NK1.1-APC-eFluor780 (clone PK136, eBioscience), anti-NK1.1-BV650 (clone PK136, Sony), anti-NKp46-BV711 (clone 29A1.4, Biolegend), anti-NKp46-V450 (clone 29A1.4, BD Horizon), anti-Sca-1-AlexaFluor700 (clone D7, Biolegend), anti-Sca-1-BV510 (clone D7, Sony), anti-S1PR5-PE (clone 1196A, Bio-Techne, Abingdon, UK), anti-TCRαβ-biotin (clone H57 597, Sony), anti-TCRγδ-biotin (clone GL3, BD Pharmingen), anti-TER119-biotin (clone TER-119, Sony) and streptavidin-PE-Cyanine5 (BD Pharmingen) for surface antigens; and anti-Eomes-PE-Cyanine7 (clone Dan11mag, eBioscience) and anti-Tbet-eFluor660 (clone eBio4B10, eBioscience) for intracellular staining.

Techniques: Isolation, Flow Cytometry, Expressing, One-tailed Test

Journal: eLife

Article Title: Haploinsufficiency of the essential gene Rps12 causes defects in erythropoiesis and hematopoietic stem cell maintenance

doi: 10.7554/eLife.69322

Figure Lengend Snippet:

Article Snippet: Antibody , APC Rat Anti-Mouse monoclonal cKit , BD Biosciences , Cat #553356 , Clone:2B8 (1 µl per 10 7 cells).

Techniques: Generated, Staining, Sequencing, DNA Extraction, Western Blot, Isolation, SYBR Green Assay

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Induced Cardiac Progenitor Cells Repopulate Decellularized Mouse Heart Scaffolds and Differentiate to Generate Cardiac Tissue

doi: 10.1016/j.bbamcr.2019.118559

Figure Lengend Snippet: (A) Schematic timeline of experimental design for reprogramming of adult wild type lung fibroblasts (derived from C57BL/6J mouse) and cardiac fibroblasts homozygous for the rtTA transgene (derived from R26-M2rtTA knock-in mouse). (B) Adult mouse lung and rtTA-cardiac fibroblasts were infected with a doxycycline inducible GFP expressing lentivirus that also contained an IRES-mCherry cassette, and greater than 90% of infected fibroblasts showed GFP/mCherry expression after doxycycline induction indicating a robustly functional expression system. (C) rtTA-cardiac fibroblasts were infected with 5 cardiac factors (MTGNB) and cultured in iCPC induction medium containing doxycycline, LIF and BIO. Cells undergoing reprogramming appeared morphologically different from fibroblasts and could be identified as small proliferative clusters after 2 weeks. (D) 4 weeks after induction of reprogramming of rtTA-cardiac fibroblasts, proliferative reporterless iCPC colonies were observed. (E) Number of reporterless iCPC colonies, derived from lung fibroblasts or rtTA-cardiac fibroblasts, observed 4 weeks after infection with 5 cardiac factors (50,000 starting fibroblasts) (n=3, SEM). (F) Reporterless iCPCs, derived from wild type lung fibroblasts and rtTA-cardiac fibroblasts, could be stably expanded without exogenous expression of cardiac factors. (G) Reporterless iCPCs expressed transcription factors Gata4, Nkx 2.5, and Irx4 which are associated with cardiac progenitor cells. (H) Flow cytometry revealed reporterless iCPC generated from rtTA-cardiac fibroblasts expressed Cxcr4, showed dim expression for Flk1, but only a small fraction was ckit positive.

Article Snippet: Flow cytometry analysis was done using cell surface markers, including Cxcr4(BioLegend 647/APC conjugate – 1:50), Flk1 (BD Pharmingen PE conjugate – 1:20), cKit (BD Pharmingen PE/Cy7 conjugate – 1:50), and PDGFRα (Santacruz PEconjugate – 1:20).

Techniques: Derivative Assay, Knock-In, Infection, Expressing, Functional Assay, Cell Culture, Stable Transfection, Flow Cytometry, Generated