CHMP2A Antibody - #DF12147

Figure 3 Poly-aneuploid RTP cells produced proliferating progenies via a viral budding-like cell division. A) Time-lapse sequences illustrating budding of an HCT116-RTP cell expressing H2B-EGFP following 8 Gy. A daughter cell (pink triangle) is being budded off. Scale bars, 25 µm. B) Representative iFISH images of HCT116 cultivated in suspension illustrating the alteration of ploidy number at indicated time points following 10 Gy. Ploidy number was visualized by CEP8 probe (human chromosome 8), DNA and cytokeratin was stained by DAPI (blue) and CK18 (golden). Scale bars, 25 µm. C) Copy numbers of chromosome 8 (CEP8, red) and centrosome (γ-tubulin, green) were detected by iFISH in unirradiated and 8Gy-treated HCT116. (Middle): A RTP cell (yellow triangle) in process of budding is surrounded by diploid progeny cells (white triangle), which is centrosome (red triangle) independent. (Right): A repopulated tumor cell is undergoing mitosis. D) Immunofluorescent staining of budding RTP cells by EdU (red), DAPI (blue) and senescence-associated β-gal (green). E) Schematic of the naming principle and cell count of cells samples collected for SMART RNA-seq. Single cells from untreated HCT116 (both p53+/+ and p53−/−) cells were defined as [N], while cells derived from pre-budding RTP cells, budding RTP cells, budded and repopulated small cells were, respectively, defined as [P], [B], [S], and [R]. F) Gene set enrichment analysis (GSEA) showing positive enrichment for budding and maturation of human immunodeficiency virus (HIV) virion in budding RTP cells relative to pre-budding RTP cells. G) Expression of ESCRT genes CHMP2A and VPS28 in violin plots and in immunofluorescent staining. Scale bar, 25 mm. iFISH: immunofluorescence in situ hybridization; ESCRT: endosomal sorting complex required for transport.