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Fig. 2 | Virology Journal

Fig. 2

From: Human papillomavirus type 16 E6 and E7 oncoproteins interact with the nuclear p53-binding protein 1 in an in vitro reconstructed 3D epithelium: new insights for the virus-induced DNA damage response

Fig. 2

In vitro 3D epithelial cultures. HPV16 DNA (FISH) staining and immunofluorescent detection of BrdU, E6 and E7 of control normal and E6E7 HPV16 infected reconstructed in vitro 3D epithelial cultures. Viral genome amplification positivity is evidenced in red in the mid and upper strata of the infected epithelium only (A, B). To show cell nuclei, sections are counterstained with DAPI (blue). S phase cells positivity to the BrdU marker is limited to the basal and basal/parabasal strata of normal (C) and infected epithelia (D) respectively. Direct analysis of E6 and E7 proteins in normal uninfected (E and G for E6 and E7 respectively) and E6E7 HPV16 positive culture sections (F-F′ for E6 and H-H′ for E7). Positive cells are shown in red, while cells nuclei are counterstained in blue with DAPI. Bar scales in 20x fields = 100 μm. Immunoblotting. Western blot analysis of E6, E7 and tubulin protein extracts derived by peeling off the epithelial layers of 3D reconstructed epithelial cultures (control normal and E6E7HPV16) from the collagen-based dermis. After run on polyacrylamide gels, proteins were blotted onto nitrocellulose or PVDF membranes (I)

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