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Fig. 8 | Stem Cell Research & Therapy

Fig. 8

From: Rapamycin regulates autophagy and cell adhesion in induced pluripotent stem cells

Fig. 8

Rapamycin treatment accelerates differentiation of human iPSCs in association with decreased NANOG expression. a Proteins were extracted from rapamycin-induced spheres and control EBs, and blotted with antibodies against LC3B, AFP, ASM, TUJ1, and β-actin. 1, 2, 3 in panel (a) indicate source of proteins from three independent iPSC lines. b Relative protein expression was quantified against β-actin and (c) compared between two methods, presented as mean ± SEM. Note a 2–3-fold increase of LC3B, AFP, ASM, and TUJ1 expression in rapamycin-induced spheres. Rapamycin-induced spheres and control EBs were plated for spontaneous differentiation for 3 days (j, k) or 5 days (di), and stained for NESTIN (k), TUJ1 (d, g), ASM (e, h), or AFP (f, i). Note the presence of more immune-positive cells in derivatives of rapamycin-induced spheres (gi) compared with cells differentiated from conventional EBs (d, f). (l) Proteins were extracted from iPSCs treated with rapamycin at 0, 1, 10, 100, 200, or 300 nM for 4 days, and immunoblotted with anti-NANOG, showing concentration-dependent reduction of NANOG expression after rapamycin treatment. m Relative abundance of NANOG expression after 4 days of rapamycin treatment. Bar = 100 μm di; 50 μm j, k. *p < 0.05, **p < 0.01. Con control, EB embryoid body

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