Additional file 2 of Tunneling nanotubes induced by pseudorabies virus facilitate viral transmission in neuronal cells to evade the immune system

Supplementary Material 2: S1 Video. Formation of filamentous structures in SK-N-SH cells infected with the PRV hSD-1/2019 strain. SK-N-SH cells were infected with the hSD-1/2019 strain at an MOI of 0.1. After 12 h, the cells were analyzed using an inverted fluorescence microscope (EVOS FL Auto). The movie consists of 120 frames captured over 10 h (1 frame/5 min) and shows the formation of filaments in SK-N-SH cells. S2 Video. The long filaments in SK-N-SH cells infected with the PRV hSD-1/2019 strain maintain stable existence. SK-N-SH cells were infected with the hSD-1/2019 strain at an MOI of 0.1. After 24 h, the cells were analyzed using an inverted fluorescence microscope (EVOS FL Auto). The movie consists of 11 frames captured over 150 min (1 frame/15 min) and shows the formation of filaments in SK-N-SH cells. S3 Video. Formation of filamentous structures in SK-N-SH cells transfected with PRV-US3 plasmids. SK-N-SH cells were transfected with the Ea-US3 plasmid. After 24 h, the cells were analyzed using an inverted fluorescence microscope (EVOS FL Auto). The movie consists of 30 frames captured over 5 h (1 frame/10 min) and shows the formation of filaments in SK-N-SH cells. S4 and S5 Videos. Transport of EGFP through TNTs induced by hSD-US3 (S3 Video) and Ea-US3 (S4 Video) plasmids transfection. Plasmids containing viral US3 and EGFP were co-transfected in SK-N-SH cells, respectively. After 24 h, the cells were analyzed using an inverted fluorescence microscope (EVOS FL Auto). The movie in S4 video consists of 30 frames captured over 5 h (1 frame/10 min) and the movie in S5 video consists of 61 frames captured over 10 h (1 frame/10 min), showing that US3-induced TNTs mediate the transport of EGFP between cells. S6 Video. Transport of virions through the tubular structures of TNTs. SK-N-SH cells were infected with hSD-mCherry virions at an MOI of 0.1. After 24 h, the cells were analyzed using a confocal microscope (ZEISS LSM 800). The movie consists of 40 frames captured over 200 min (1 frame/5 min) and shows mCherry-labeled virions being transmitted between cells through the tubular structures of TNTs. S7 Video. Intercellular transport of multiple vesicular structures within TNTs. SK-N-SH cells were infected with hSD-mCherry virions at an MOI of 0.1. After 24 h, the cells were analyzed using an inverted fluorescence microscope (EVOS FL Auto). The movie consists of 68 frames captured over 340 min (1 frame/5 min) and shows multiple vesicular structures being rapidly transported between cells within TNTs. S8 Video. Transport of virions through vesicular structures within TNTs. SK-N-SH cells were infected with hSD-mCherry virions at an MOI of 0.1. After 24 h, the cells were analyzed using a confocal microscope (ZEISS LSM 800). The movie consists of 25 frames captured over 120 min (1 frame/5 min) and shows mCherry-labeled virions being transmitted via the movement of vesicular structures within TNTs.