Tcs sp8 sted x laser scanning

Wild type and WDR77^-/- HEK293T cells were seeded in 6-well dish with a density of 60%. 12 h post-infection with VSV, fluorescent images were taken for live cells using Olympus IX71 inverted fluorescence microscope. To determine subcellular localization of WDR77, HEK293T cells were transfected with pcDNA3-HA-WDR77 and Flag-MAVS by Lipofectamine 3000. Twenty-four hours after transfection, cells were infected with or without SeV for 8 h. Cells were then washed with pre-warmed 1×PBS, fixed with 4% formaldehyde for 15 min, and permeabilized with 0.5% Triton X-100 for 15 min. After blocking with 3% bovine serum albumin (Roche,10735078001) at room temperature for 1 h, the cells were incubated with anti-HA or anti-Flag antibodies respectively overnight at 4 °C. After incubation, cells were washed with pre-warmed 1×PBS three times. Before fluorescent imaging, cells were incubated with secondary antibodies as indicated and 1 μg/ml DAPI (Beyotime, C1002). All the images were taken with Leica TCS SP8 STED X laser scanning confocal microscope.

Wild type and WDR77^-/- HEK293T cells were seeded in 6-well dish with a density of 60%. 12 h post-infection with VSV, fluorescent images were taken for live cells using Olympus IX71 inverted fluorescence microscope. To determine subcellular localization of WDR77, HEK293T cells were transfected with pcDNA3-HA-WDR77 and Flag-MAVS by Lipofectamine 3000. Twenty-four hours after transfection, cells were infected with or without SeV for 8 h. Cells were then washed with pre-warmed 1×PBS, fixed with 4% formaldehyde for 15 min, and permeabilized with 0.5% Triton X-100 for 15 min. After blocking with 3% bovine serum albumin (Roche,10735078001) at room temperature for 1 h, the cells were incubated with anti-HA or anti-Flag antibodies respectively overnight at 4 °C. After incubation, cells were washed with pre-warmed 1×PBS three times. Before fluorescent imaging, cells were incubated with secondary antibodies as indicated and 1 μg/ml DAPI (Beyotime, C1002). All the images were taken with Leica TCS SP8 STED X laser scanning confocal microscope.

Wild type and WDR77^-/- HEK293T cells were seeded in 6-well dish with a density of 60%. 12 h post-infection with VSV, fluorescent images were taken for live cells using Olympus IX71 inverted fluorescence microscope. To determine subcellular localization of WDR77, HEK293T cells were transfected with pcDNA3-HA-WDR77 and Flag-MAVS by Lipofectamine 3000. Twenty-four hours after transfection, cells were infected with or without SeV for 8 h. Cells were then washed with pre-warmed 1×PBS, fixed with 4% formaldehyde for 15 min, and permeabilized with 0.5% Triton X-100 for 15 min. After blocking with 3% bovine serum albumin (Roche,10735078001) at room temperature for 1 h, the cells were incubated with anti-HA or anti-Flag antibodies respectively overnight at 4 °C. After incubation, cells were washed with pre-warmed 1×PBS three times. Before fluorescent imaging, cells were incubated with secondary antibodies as indicated and 1 μg/ml DAPI (Beyotime, C1002). All the images were taken with Leica TCS SP8 STED X laser scanning confocal microscope.