Eight well cell culture slide

Cells were seeded on an eight-well cell culture slide (SPL, Korea) at a density of 3 × 10 5 cells per well, and each well was infected with 20 µl of Mr-IAG lentivirus solution. Four days post-transduction, the medium was discarded, and cells were washed twice with crustacean physiological saline, and fixed with 4% paraformaldehyde for 15 min at RT. Cells were blocked and permeabilized with blocking solution containing 2% normal goat serum, 0.1% Triton X-100, 0.05% Tween-20 in PBS for 1 h at RT. Thereafter, cells were incubated overnight at 4 • C, in a humidity chamber, with rabbit anti-Mr-IAG antibody, as previously described and validated (Ventura et al., 2011) . The primary antibody was diluted to 1:200 in blocking solution. After three washes with PBS, the cells were incubated for 1 h at RT with donkey anti-rabbit IgG Alexa Fluor-546 (Thermo Fisher Scientific, United States)
diluted to 1:500 in PBS containing 0.2% fish skin gelatin (Sigma, Germany). Non-specific staining was performed with blocking solution without primary antibody. Following three washes with PBS, slides were mounted with Fluoromount-G R containing DAPI (Southern Biotech, United States) and examined under an Olympus Fluoview FV1000 laser scanning confocal microscope.