Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Forming megakaryocytes from murine induced pluripotent stem cells by the inducible overexpression of supporting factors

doi: 10.1002/rth2.12453

Figure Lengend Snippet: (MK) differentiation from R26M2K2 iPSCs. (A) Differentiation scheme for MK and platelet generation from murine iPSCs. After retroviral transduction, iPSCs were differentiated into EBs by orbital shaking for 6 days (day −6); at day 5, hematopoietic differentiation was enforced by addition of SCF and interleukin‐3. At day 1, CD41 positive early hematopoietic progenitors were selected by magnetic‐associated cell sorting and seeded onto OP9 feeder cells with thrombopoietin and stem cell factor and induced by doxycycline. After day 14, MK morphology and surface marker expression were analyzed. (B) CD41 expression on early hematopoietic progenitors after EB dissociation. (n = 3‐9, mean ± SD, no differences by one‐way analysis of variance ANOVA with Dunnett’s multiple comparison test). (C) Representative flow cytometry plots of cells after embryoid body (EB) dissociation showing expression of CD41 that is not coexpressed with the pluripotency marker Stage‐specific embryonic antigen 1 (left plot). No GFP expression could be detected at that time, (right plot). (D) Green fluorescent protein (GFP) expression in EBs measured at the day of dissociation in the uninduced state. GFP expression in MKs after 2‐week induction with doxycycline. No expression after 1‐week withdrawal of doxycycline during further cultivation of MKs (GATA‐1 transduced MKs are shown, mean ± SD, n = 3‐5) (E) Percentage of CD41 expression and CD41/CD42d double positive cells at days 14 of differentiation on hematopoietic cell differentiated on OP9 feeders, (n = 3‐8, mean ± SD, one‐way ANOVA with Dunnett’s multiple comparison test, *** P < .00; Mann‐Whitney test, * P = .006). (F) Ratio of CD42d to CD41 expression as a measure of differentiation. (G) Quantification of CD41 + MK at day 14 ((F) and (G): n = 3‐8, mean ± SD, Mann‐Whitney test)

Article Snippet: At day 5 of EB formation, medium was supplemented with 10 ng/mL murine interleukin‐3 and 30 ng/mL murine stem cell factor (SCF) (PeproTech, Rocky Hill, NJ, USA).

Techniques: Transduction, FACS, Marker, Expressing, Flow Cytometry, MANN-WHITNEY

Journal: Cell stem cell

Article Title: Inhibition of Inflammatory Signaling in Tet2 Mutant Preleukemic Cells Mitigates Stress Induced Abnormalities and Clonal Hematopoiesis

doi: 10.1016/j.stem.2018.10.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: mSCF peptide , PeproTech , #250–03.

Techniques: Multiplex Assay, Cell Isolation, Reverse Transcription, SYBR Green Assay, Chromatin Immunoprecipitation, DNA Purification, Recombinant, Lysis, Western Blot, Flow Cytometry, Software

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: Chemical compound, drug , mSCF , Peprotech , Cat # 250–03 , Cytokine.

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