Fig. 3

Effect of NAMPT or SIRT2 inhibition on the iPS cell pluripotency. a Analysis of the expression of the pluripotent stem cell surface markers, SSEA4 and Tral-1-60, on FK866 (left) or AC93253 (right)-treated human iPS cells using FACS. DMSO was used as vehicle control. Data represent means ± SD from two independent experiments, each in triplicates. b mRNA expression of pluripotency genes in human iPS cells treated with FK866, AC93253, or DMSO for 48 h. Fold changes of pluripotency markers relative to DMSO-treated cells are shown. Data represent means ± SD from two independent experiments, each in triplicates (*p < 0.05, **p < 0.01, ****p < 0.0001). c mRNA expression of genes specific for the three germ layers in human iPS cells treated with FK866, AC93253, or DMSO for 48 h. Fold changes of markers of differentiation relative to DMSO-treated cells are shown. Data represent means ± SD from two independent experiments, each in triplicates (*p < 0.05, **p < 0.01, ****p < 0.0001). d iPS cells were treated with DMSO, 1 nM of FK866, or 50 nM of AC93253 for 48 h with subsequent ICH analysis of the ectodermal and endodermal differentiation, as described in “Material and methods”. Representative images are depicted, no Ab: control staining w/o antibody. e Ectodermal and endodermal differentiation of iPS cells in the presence of DMSO, 1 nM FK866, or 50 nM AC93253 was evaluated, as described in “Material and methods”. Representative images are depicted, no Ab: control staining w/o antibody. f iPS cells were treated with DMSO, 1 nM FK866, or 50 nM AC93253 for 48 h, after that ectodermal and endodermal differentiation was performed, as described in “Material and methods”, without addition of inhibitors. Representative images are depicted, no Ab: control staining w/o antibody