Figure S3 from Epstein–Barr Virus miRNA BART2-5p Promotes Metastasis of Nasopharyngeal Carcinoma by Suppressing RND3

<p>Supplementary Figure 3 (a) CNE2 were transiently transfected with control mimic (Cel- 39), BART2-5p mimic, BART2-5p mimic + pcDNA3.1Vec, BART2-5p mimic + RND3 or BART2-5p and Y-27632 treatment. For control cells and BART2-5p transfected cells, F-Actin staining was performed to visualize stress fiber formation at 6 h after transfection. For cells transfected by BART2-5p and pcDNA3.1Vec or RND3 expressing vector, vector or RND3 was transfected 24 h earlier than transfection of BART2-5p to allow gene expression in cells. Pictures were taken at 6 h after transfection of BART2-5p. 1μM ROCK inhibitor Y-27632 was added to culture medium for a 15min incubation at 6 h after transfection of BART2-5p before fixation. (b) CNE2 cells with BART2 expression or RND3 knock down were lysed for Western Blot. Either BART2 overexpression or RND3 knock down led to increased phosphorylated MLC2 and MYPT1. (c) Western Blot shows induced activation of ROCK signaling by BART2 in CNE2 cells could be abolished by reinstatement of RND3 or Y-27632 treatment. (d) Images of Western Blot show alteration of key components in ROCK signaling pathway in siBART2-5p transfected or RND3 overexpressed HONE-1 EBV (+/-) cells.</p>