The invasion of tobacco mosaic virus RNA induces endoplasmic reticulum stress-related autophagy in HeLa cells

Li, Li; Wang, Li; Xiao, Ruijing; Zhu, Guoguo; Li, Yan; Liu, Changxuan; Yang, Ru; Tang, Zhiqing; Li, Jie; Huang, Wei; Chen, Lang; Zheng, Xiaoling; He, Yuling; Tan, Jinquan

To observe the location of TMV-RNA, autophagosomes and ER, 4 h post-transfected live cells were detected on a Zeiss LSM 510 Inverted Live-Cell Confocal System following staining with ER-Tracker TM Blue-White DPX (Invitrogen). The treated cell dish was placed on the stage of a temperature-controlled microscope. Unless otherwise noted, experiments were conducted at 37 • C. CTAB-QD-labelled TMV-RNA, GFP-LC3-labelled autophagosomes and ER were detected by using a filter set consisting of a D565/500/435 nm exciter, a D615/530/485 nm emitter. At the transfection time point (4 h), TMV-RNA showed up as red dots and blue fluorescence represented ER. The formation of GFP-LC3-labelled structures (GFP-LC3 'dots') represent autophagosomes. Notably, the red dots were co-located with some punctate GFP-LC3 staining, meanwhile just appeared on ER (blue), whereas no red dot was found, as well as GFP-LC3 'dots' decreased in cells transfected with GFP-LC3 alone. All these showed that TMV-RNA co-located with autophagosome on ER when it was transfected into HeLa cells for 4 h.