<p>(<b><i>A</i></b>) Morphological changes in culture during differentiation of HTCE cells. (<b><i>B</i></b>) AE3 and AE1 keratin panel staining in HTCE cell differentiation. (<b><i>C</i></b>) Time course of K12 mRNA expression following induced differentiation of HTCE cells. Differentiation of HTCE cells was induced by adding 1.2 mM calcium and 5% FBS in the normal culture condition up to 120 h. (<b><i>D</i></b>) Detection of K12 and p63 expression in HTCE cell differentiation by Western blot up to 120 h. (<b><i>E</i></b>) Expression of K12 and p63 mRNAs and proteins in human limbal stem/progenitor (HLS/P) and corneal epithelial (HCE) cells. Cells were grown and induced to differentiation in chamber slides and photos were taken by light microscopy. These cells were then washed in PBS and fixed with 95% ethanol before staining. Staining protocols were used as suggested by manufactures. Furthermore, RNAs were extracted from HTCE, HLS/P and HCE cells, and reverse-transcribed into cDNAs before real-time qPCR analysis was performed using K12-specific primers as indicated in Materials and Methods. Symbol “*” indicates significant differences after 72 h induction (<i>p</i><0.05, n = 6). Photos were taken by a Zeiss AXIO microscope (x20).</p
