Fig. 4

NAMPT or SIRT2 inhibition suppresses hematopoietic differentiation of iPS cells. a Work flow of EB-based hematopoietic differentiation of iPS cells in the presence of DMSO, FK866, or AC93253. b Representative images of differentiated iPS cells treated with indicated drugs for 25 days. Images were taken using a Nikon Eclipse TS 100 microscope. Scale bars: 50 μm. c Number of viable hematopoietic cells generated in the presence of FK866 or DMSO using EB-based hematopoietic differentiation protocol at indicated time points, as assessed using trypan blue dead cell exclusion assay. Fold changes relative to DMSO-treated cells are shown. Data represent means ± SD from two independent experiments, each in duplicates (**p < 0.01). d Flow cytometry analysis of suspension cells harvested from EBs culture on day 18 and day 25 of differentiation for iPS cells treated with DMSO or FK866. Fold change difference of the absolute numbers of each cell fraction relative to DMSO are shown. Data represent means ± SD from two independent experiments, each in duplicates (*p < 0.05, **p < 0.01, ***p < 0.001). e Representative cytospin images of suspension cells derived from differentiated human iPS cells treated with DMSO or FK866 on day 25 of culture. Scale bars: 400 μm