Expression of siPARI-1-resistant eGFP-PARI rescues the ICRF-193-induced midbody lifetime increase. (A) Left panel: WT siPARI-1 target sequence, features highlighting seed sequences. Right panel: siPARI-1–resistant PARI sequence; features show where three silent point mutations were introduced by cloning. (B) RT-qPCR measures the mRNA levels of PARI in HeLa cells after transfecting twice with 25 nM siCtrl, siPARI-1, and siPARI-2. Relative mRNA levels have been normalized to siCtrl. One-sample t and Wilcoxon test (mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, N = 4). (C) Cells treated with ICRF-193 progressing from anaphase to midbody formation (SiR-tubulin) and its disassembly. Cells have been synchronized and treated as illustrated in Fig. 5. Open arrowhead specifies midbody disassembly. Scale bar: 5 μm. (D) SuperPlot showing the quantification of the midbody lifetime (time from midbody formation to disassembly). Student’s paired t test (mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n = 165, N = 4).