Fig 5. Localisation and expression pattern of E6/E7 and its targets during HPV11 productive lifecycle in condyloma acuminatum.

(A) H&E staining of condyloma acuminatum biopsy with enlargement areas of lesion (left) and non-infected epithelium (right). (B-D) E6/E7 RNAScope, MCM7 and K10 immunofluorescence staining were carried out on adjacent sections. Nuclei were counterstained with DAPI (blue). Scale = 200 μm. Enlargement images are shown at the bottom, lower left is the lesion and lower right is the non-infected area. The dotted lines indicate the position of the basal layer. (E-F) E6AP and NHERF1 protein expression in infected (lower left) and non-infected (lower right) region of the biopsy. Scale = 200 μm. (G-I) The graphs show quantifications of basal cell density (nuclei per mm) by counting cells per 100 μm distance along the basal layers using Image J (G), % MCM positive cells in the basal layer (H), and distance from the basal lamina to the bottom of K10-positive cells (μm) (I). (J-K) show quantifications of E6AP and NHERF1 protein expression in the basal layer for HPV-infected and non-infected tissues. Integrated intensity (sum of the pixel intensity) of E6AP or NHERF1 stain in the basal layers of infected and non-infected tissues was measured using ImageJ. At least three different tissue samples and three representative images from each sample were included in the quantification. Graphs show mean values ± standard errors of amongst different condyloma biopsies. P values were calculated with student t tests. *, P ≤0.05; **, P ≤0.01; ****, P ≤0.0001.