Overexpression of CaSR blocks acquisition of stem cell-like characteristics. HT29 cells overexpressing the CaSR (HT29CaSR) or empty transfected cells (HT29EMP) were grown in stem cell media. (A) Ectopic expression of CaSR blocked acquisition of the pluripotency markers Nanog, Oct3/4, Stella, and FOXC2. (B) Treatment with NPS R-568 or ovexpression of CaSR diminished expression of pluripotency-associated markers Sox2, Nanog, Oct4 and CD44. (C) Extreme limiting dilution assay revealed a significantly lower sphere forming ability of stem-like cells in HT29CaSR cells. Data represent mean ± SEM of 3-5 independent experiments. Scale bar: 50 μm. Statistical significance was calculated using t test. *p < 0.05, **p < 0.01.