MRCKα 1–478 cleavage product induces iEAAR assembly and epithelial extrusion. (A) Transfection of mCherry-MRCKα 1–478 in MDCK, HeLa, A431, and Caco2 cells triggers iEAAR assembly. (B) Time-lapse sequence of a mCherry-MRCKα 1–478 expressing MCF10A cell undergoing the process of extrusion from a confluent epithelium. (C) Fate of MCF10A cells expressing mCherry alone, mCherry-MRCKα 1–478, or mCherry-MRCKα 1–478 KD. Cells were assigned to two categories: those that remained alive in the epithelium (Adherent) and those that were removed from the epithelium (Extruded). (D) MCF10A cells expressing mCherry-MRCKα 1–478 and show the presence of iEAAR despite the treatment with tBid inhibitor and the caspase inhibitor zVAD-FMK. (E) Analysis of the percentage of MCF10A cells transduced with mCherry-MRCKα 1–478 showing iEAAR in the presence of different concentrations of zVAD-FMK. (F) Treatment of MCF10A cells with tBid inhibitor or zVAD-FMK does not prevent epithelial extrusion induced in MCF10A by mCherry-MRCKα 1–478 transfection. (G) The expression of mCherry-MRCKα 1–478 in MCF10A induces the formation of iEAAR without the induction of apoptosis (negative for active caspase 3 staining).