<p>Palmitoylation of the SVCV G protein is essential for viral budding.</p>

Scale bars,10 μm. <b>(F)</b> ZF4 cells expressing EGFP-SVCV G were treated with control solvent or 2-BP (50 μM) for 24 h at 16°C. Cell membranes were stained with Dil and nuclei with DAPI. Scale bars,10 μm. <b>(G)</b> ZF4 cells were transfected with EGFP-SVCV G or the mutants for 24 h at 16°C. Cell membranes were stained with Dil and nuclei with DAPI. Scale bars,10 μm. <b>(H)</b> ZF4 cells were co-transfected with SVCV G and SVCV M for 24 h at 16°C and 28 °C. The protein levels of SVCV-M and SVCV-G in the cell lysate and culture supernatants were measured by Western blotting. <b>(I)</b> ZF4 cells were transfected with plasmids expressing SVCV G/the mutants and SVCV M at 16 °C and subsequently mock treated (SVCV G/the mutants and SVCV M groups) or treated with 50 mM 2-BP (SVCV G 2-BP group).

The protein levels of SVCV-M and SVCV-G in the cell lysate and culture supernatants were measured by Western blotting. <b>(J)</b> ZF4 cells were infected with SVCV (MOI = 10) under different temperature conditions. Following 12 hours post-infection, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy to observe SVCV budding. Representative images are shown in (J), with scale bars of 5 μm and 1 μm. <b>(K)</b> The budding efficiency of SVCV in ZF4 cells cultured at different temperatures was assessed by calculating the ratio of intracellular and extracellular infectious virus titers to the total infectious titer at 12 hours post-infection.

Data was presented as mean ± SD from three independent experiments. Statistical analysis was performed using Student’s <i>t</i>-test between different groups and the asterisk (*) indicates significant differences between groups*<i>p</i> < 0.05, **<i>p</i> < 0.01, ***<i>p</i> < 0.001.</p>