Removal of the midbody remnant greatly interferes with PC formation in MDCK cells. (A) Representative images of the TUSP procedure. DIC, differential interference contrast; (B and C) TUSP was used to remove the midbody remnant (B, bottom). As a control (Ctrl), TUSP was applied to cells with a remnant in a zone of the plasma membrane distant from it (B, top). After 24 h, the same cells were examined for the presence of a PC. The green and white arrowheads mark the midbody remnant and the PC, respectively. The circles indicate the plasma membrane zone subjected to TUSP. Bars, 3 µm. (C) The percentage of ciliated cells was quantified 24 h post-TUSP. Data represent the mean + SEM from six independent experiments (n = 26 control cells and 27 cells whose midbody remnant was removed; χ2 test). (D) The distribution of podocalyxin, ZO-1, and GFP-tubulin was analyzed in cells in which the remnant was removed by TUSP. The arrowheads point to the midbody remnants, and the circles mark the plasma membrane zone subjected to TUSP. Bars, 5 µm.
