Fig 6.

Confocal microscopy of ISKNV internalization into MFF-1 cells. Cells grown on coverslips were prechilled and then infected with ISKNV at 4°C for 1 h. Noninternalized viruses were washed up by using TBS and fixed with methanol at 0 h (B), 0.5 h (C), 1 h (D), 2 h (E), and 4 h (F), while uninfected cells (A) were used as a negative control. The cells were analyzed by IFA using anti-mCav-1 and anti-ORF101L antibodies and were then viewed under a confocal microscope (LSM510; Zeiss) equipped with 555/488-nm argon-krypton and 543-nm helium-neon lasers. ISKNV particles (red fluorescence), mandarin fish caveolin-1 (green fluorescence), and nuclei (blue fluorescence) were visualized with Hoechst 33342. Magnification, ×600.