Fig. 3

Prolonged expression of HAS1 increases clonal diversity. a A spectrum of clonal diversities was observed in the Dox-induced populations (lower panels) varying widely in nuclear/cytoplasmic sizes and shapes. The Dox induction was verified with higher GFP expression in +Dox cells (lower panel) then the –Dox controls (upper panel). This figure is representative of at least 5 individual induction experiments. b MCF10A cells were transfected with plasmids as indicated in Fig. 1a. HAS1 transfected cells show variation of the nuclear size and shapes demonstrating clonal variations. The resulted cell population were found homogenous in control mock transfected (upper left) as well as irrelevant control protein expressing (LMA2) cells (upper right). White arrows show micro-nucleus, asterisk indicate multi-nucleated or huge donut-shaped nucleus, and the red bar is 50 μm. c DLD1 colorectal adenocarcinoma cell line was transfected and selected with Tet-inducible HAS1 or HAS2 or control (pTET) systems, fixed and stained the nucleus with DAPI (blue). HAS1 and HAS2 cells showed much diversity in nuclear perimeter size in comparison to pTET cells (control). The cells were observed under confocal microscope and photographed for multiple fields from each slides. The nuclear perimeter (without micronucleus) were marked and measured with Olympus FV1000 software. The average perimeter is marked with thick horizontal bars. The range of nuclear size/morphological diversity is shown in dashed elliptical areas