Hyperosmolar sorbitol induces Eomes in TSCs in a MAPK8/9-dependent (MAPK8/9-inhibitor sensitive) manner. TSCs were incubated with 400mM sorbitol for 0–1hr ± MAPK8/9 inhibitors LJNKl1 or SP600125 (data not shown), fixed, and stained for Eomes. TSCs were cultured for 1hr in media alone (A, B), with 400mM sorbitol (C, D), with 400mM sorbitol and MAPK8/9 inhibitor LJNKl1 (E, F), or LJNKl1 alone (G, H), and stained for Eomes. TSCs were incubate with 400mM sorbitol but developed without anti-Eomes antibody. The micrographs in (B, D, F, H, and J) are Hoechst-stained nuclei corresponding to the cells in (A, C, E, G, and I), respectively. The histogram shows mean fluorescence intensity of 6 replicates of the panels shown in A–I as indicated. Error flags are SD of the means for the 6 replicates. Statistical analysis by ANOVA, with Duncan post hoc test, compares the following pairs; (a) C to A (p=0.03), (b) E to C (p=0.004), (c) E to A (p=0.48), and (d) G to A (p=0.7).