Fig 5.

Mitosis is associated with the decay process in dual-replicon cells. (A) Validation of the fluorescent hGem probe for cells in S/G₂/M phases of the cell cycle. 7.5-GFP-hGem cells were stained with PI and analyzed for GFP and PI by flow cytometry. As shown, the GFP⁺ cells are highly enriched in the S/G₂/M phases of the cell cycle, while the GFP⁻ cells are enriched for the G₁ phase of the cell cycle. (B) Dual-replicon cells are equally competent as single-replicon cells for cell cycle progression. 7.5-GFP-hGem cells were transfected with Jc1/Δ^GFP and Jc1/Δ^mCherry RNA. Dual-replicon (EBFP2⁺/mCherry⁺) and single-replicon (mCherry⁺ only) cells were isolated in G₁ (GFP⁻) and S/G₂/M (GFP⁺) phases of the cell cycle by FACS. No difference was observed by Student's t test in the cell cycle phase of single- and dual-replicon cells at any time point (P > 0.2, n = 3). (C) As dual-replicon cells progress through mitosis, single-replicon cells are enriched. 7.5-GFP-hGem cells were transfected with Jc1/Δ^EBFP2 and Jc1/Δ^mCherry. Dual-replicon (EBFP2⁺/mCherry⁺) S/G₂/M-phase (GFP⁺) cells were isolated by FACS and analyzed by flow cytometry for the dual- to single-replicon transition, as well as cell cycle progression. Error bars indicate +SEM (n = 3).