Supporting information related to the tumor cell-selective E4orf4 phenotypes in cell lines of various origins.(A) Representative epifluorescence images of cells from various origins transduced with Ad-RFP or Ad-E4orf4-mCherry, showing transduction efficiencies 48 h after transduction, associated with the absence (nontumorigenic cells: HaCaT and hTERT RPE-1) or presence (cancer cell lines: MDA-MB-468, MDA-MB-231, U2OS, HeLa, and 22Rv1) of morphological changes (cell rounding and blebbing). Bars: 50 µm. (B) Quantification of the percentage of cell blebbing and nuclear condensation cell, as scored relative to cells expressing RFP/E4orf4-mCherry 24–36 h after transduction, before and after blebbistatin treatment (50 µM, 2 h), for datasets represented in C (means ± SEM [n ≥ 3]; *, P < 0.05; ****, P < 0.0001; ns, not significant. (C) Maximum intensity projections of confocal image stacks of representative RFP- or E4orf4-mCherry-expressing cells; the nuclear circularity index (C.I.) was measured from lamin A/C staining; for the 22Rv1 cell panel, the indicated circularity index corresponds to the nucleus designated by a dotted white line. ns, not significant. Bars, 10 μm.