Figure 3
MiR203 induces G0/G1 arrest and blocks colony formation capacity in a manner that is rescued by ΔNP63α. (a) Western blot analysis of protein lysates from IMEC (left) and HC11 (right) cells transiently transfected with mature miR203 (60 nm) for 48 h and probed with an anti-P63 antibody (4A4). (b) 10 × Phase contrast microscopy of HC11 cells transiently transfected with mature miR203 (60 nm) for 72 h. (c) PI cell cycle analysis was performed via flow cytometry of HC11 cells transiently transfected with mature miR203 for 72 h (cells were treated with nocodizole to synchronize cells before PI cell cycle analysis). (d) IMEC cells were either infected with an empty retrovirus or a retrovirus expressing the open reading frame of ΔNP63α (pLPC-ΔNP63αORF) and then transiently transfected with either a miR-control or mature miR203 (60 nm) and then plated for colony formation assay. Colonies were stained with crystal violet and imaged. All experiments were performed in triplicate (n=3)
