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. 2018 Nov 16;37(24):e100409. doi: 10.15252/embj.2018100409

Figure EV1. Pharmacological modulation of PKCδ activity preserves HSPC activity in vitro .

Figure EV1

  • A, B
    One hundred HSCs were sorted from WT mice and cultured in 96‐well (U‐bottom) plates in triplicate in the presence of mSCF (20 ng/ml) and mTPO (20 ng/ml) and with or without Mallotoxin (MTX, 5 μM) or Indolactam V (Indo‐V, 10 μM) for indicated time. (A) FACS histograms and (B) bar graph show the percentage of Lin‐ cells (pre‐gated on live cells).
  • C
    At the indicated time of culture, cells were analyzed for LSK phenotype. Representative FACS plots showing the percentage of cells retaining LSK phenotype. Data are representative of two independent experiments (n = 5 mice total per treatment group).
  • D
    Schematic of competitive reconstitution analysis of MTX‐treated WT HSPCs. After 13 days in culture, cells were transplanted into recipient mice along with 1 × 106 competitive total BM cells (CD45.1+). Percentage of total donor‐derived cells (CD45.2+), B cells (B220+), and myeloid cells (CD11b+ Gr1+) in the peripheral blood was analyzed at indicated time (n = 6 mice per condition).
Data information: All data shown as mean ± SEM. *P < 0.05 and **P < 0.01 by repeated measures one‐way ANOVA analysis with Bonferroni posttest.