Figure 4

Co-localization of hnRNPC with HDV antigens and RNA in human liver cells. (A-C) Double indirect immunofluorescence was used to detect HDAg and hnRNPC proteins in cultured Huh7-D12 cells. Cells were fixed with 3.7% formaldehyde, permeabilized with 0.5% Triton X-100, and incubated with an anti-HDAg antibody (A, green staining) and an anti-hnRNPC antibody (B, red staining). Combined in situ hybridization and immunofluorescence was performed to detect HDV RNA and the hnRNPC protein. After fixation and permeabilization, cells were hybridized with a digoxigenin-labeled probe to detect HDV RNA (D, green staining) and with an anti-hnRNPC antibody (E, red staining). Overlaps of images are shown in panels C and F. (G) 10 individual cells from each experiment were analyzed using the ImageJ software and the JaCoP plugin to determine the Mander's overlap coefficient.