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. 2017 Feb 28;6:e23897. doi: 10.7554/eLife.23897

Figure 2. CLCuMuV infection activates autophagy.

(A) Representative confocal microscopy images of dynamic autophagic activity revealed by specific autophagy marker CFP-NbATG8f in plants infected with CLCuMuV plus CLCuMuB (CA+β). (B) Quantification of the CFP-NbATG8f-labeled autophagic puncta per cell from (A). More than 500 mesophyll cells for each treatment were used for the quantification. Relative autophagic activity in virus infected plants was normalized to that of control plants, which was set to 1.0. Values represent means ± SE from three independent experiments. (*) p<0.05. (C) Representative TEM images of autophagic structures. Ultrastructure of autophagic bodies (arrows) was observed in the vacuoles of mesophyll cells of uninfected control and plants infected with CA+β. V for vacuole. (D) Autophagosome-like structures from (C) were quantified. At least 30 cells for each treatment were used for the quantification. Relative autophagic activity in virus infected plants was normalized to that of control plants, which was set to 1.0. Values represent means ± SE from three independent experiments. (*) p<0.05.

DOI: http://dx.doi.org/10.7554/eLife.23897.009

Figure 2.

Figure 2—figure supplement 1. Transcription pattern of autophagy-related genes were altered during CLCuMuV infection.

Figure 2—figure supplement 1.

Quantitative RT-PCR was performed using total RNA isolated from the leaves of virus infected plants and non-infected plants. Expression data relative to non-infected plants are normalized to that of NbeIF4α. Values are means ± SE from three independent experiments. (*) p<0.05.
DOI: http://dx.doi.org/10.7554/eLife.23897.010
Figure 2—figure supplement 2. Viral infection decreased NbJoka2/NBR1 protein level.

Figure 2—figure supplement 2.

(A) Western blot assays showed that NbJoka2 protein level was reduced due to the increased autophagy flux in CLCuMuV-infected plants. NbJoka2 was detected with anti-NBR1 polyclonal antibody. Stars indicate expected band size. (B) mRNA level of NbJoka2 was unchanged in CLCuMuV-infected plants. Real-time RT-PCR of NbJoka2 was used to determine mRNA level. Values represent means ± SE from three independent experiments. (*) p<0.05.
DOI: http://dx.doi.org/10.7554/eLife.23897.011