Figure 2

Myb expression during confluence/hormone-induced differentiation of HC11 cells. HC11 were proliferating (Day 1), grown to confluence (Day 2), and then differentiated by removing epidermal growth factor (EGF) for 24 hours (Day 3), followed by addition of differentiation medium containing lactogenic hormones (Days 4 to 9). (a) HC11 cells grown on glass cover slips were stained for DNA (4',6-diamidino-2-phenylindole [DAPI]; blue), lipid vesicles (Nile Red; red), and viewed under fluorescence microscopy at Day 1 and Day 9. (b) HC11 cells were grown in 24-well dishes, and induced to differentiate as described. Shown are light micrographs of confluent cells and 'domes' (arrow) which form in the presence of differentiation inducing medium. (c) Quantitative analysis of Nile Red staining by flow cytometry. HC11 cells were analyzed for lipid vesicle formation during the different stages of differentiation. (d) Quantitative PCR of MYB mRNA from HC11 cells undergoing differentiation. HC11 cells were analyzed each day during the differentiation process. Standard deviation is represented by error bars. (e) Western blot analysis of MYB protein from total cell lysates of HC11 cells during the differentiation process. A ß-tubulin loading control is also shown. Results are indicative of two independent experiments.