Fig. 2: Chronic infection of HAs with HCMV clinical isolates, the appearance of CEGBCs as well as colony formation in soft agar, and the phenotypic characterization of CEGBCs.

a HAs time-course infection with HCMV-DB and BL strains (MOI = 1). Red arrows showing the generated CEGBCs. Magnification ×100, scale bar 100 μm. Uninfected HAs were used as a control. b An inverted light microscope was used to closely follow up the chronic CEGBCs-DB and BL cultures and the appearance of several structures; magnification 200x, scale bar 100 µm. c FACS staining of Myc and EZH2 in HAs infected with HCMV-DB and BL; uninfected HAs were used as a negative control. The fold regulation of oncogenes and tumor suppressor genes (d) as well as cell cycle genes (e) was assessed in uninfected HAs and HAs infected with HCMV-DB and BL using RT² Profiler PCR Arrays. f Colony formation in soft agar seeded with CEGBCs-DB and BL (MOI = 1); UI HAs and HAs-HSV were used as controls. Formed colonies were observed under an inverted light microscope (Magnification 200x, scale bar 100 µm). g Vimentin expression by FACS and confocal microscopy in CEGBCs-DB and BL; uninfected HAs were used as a control. Nuclei were counterstained with DAPI; magnification ×63, scale bar 10 μm. h FACS staining of CD44 in CEGBCs-DB and BL. Uninfected HAs were used as controls. i The fold regulation of EMT genes was assessed in UI HAs and HAs infected with HCMV-DB and BL using RT² Profiler PCR Arrays. j Histogram depicting the expression of PN markers (OLIG2, CD133, and SOX2), MES markers (CD44, EGFR, and MET), as quantified by RT-qPCR in CEGBCs-DB and BL. **p-value ≤ 0.01. Data are represented as mean ± SD of two independent experiments.