Figure 2
Treatment with a high-dose GSK3β inhibitor efficiently induced caudal hindgut. (A) A schematic diagram of hindgut maturation after posterior DE induction. (B) An expression analysis of the hindgut markers HOXA13 and HOXD13 after four days of maturation following posterior DE induction with the indicated dose CHIR by qPCR (n = 3 independent experiments; mean ± SEM). (C) Immunostaining of HOXA13 and HOXD13 in differentiated cells. Representative fluorescence microscopic images for treatment with 1 µM (upper panels) and 5 µM CHIR (lower panels) are shown. Scale bar, 100 µm. (D) The proportions of HOXA13 (left panel)- and HOXD13 (right panel)-positive cells (n = 3 independent experiments; mean ± SD). *p < 0.05; two-tailed paired t-test. Abbreviations: iPSc, induced pluripotent stem cell; DE, definitive endoderm; CHIR, CHIR99021; BPE, bovine pituitary extract; ATRA, all-trans retinoic acid.
