CSH1 increases and ID2 decreases in 24 h after 400 mM sorbitol is removed from cultured TSCs. (A) TSCs were cultured for 24 h without or with 400 mM sorbitol, or cultured with sorbitol for 24 h followed by 24 h without sorbitol, cells were lysed, proteins were fractionated by SDS–PAGE and immunoblotted for CSH1, ID2, and reprobed for actin (ACTB) as a loading control. (B) Histograms below each immunoblot show either CSH1 or ID2 normalized to the loading control and this ratio is set to 1 for unstressed cells/embryos. Y error bars show S.D. of triplicate experiments.