Figure 3: LINE-1 RNA in P-bodies and stress granules co-localizes with autophagosomes and is likely degraded by autophagy with ORF1p. | Nature Communications

Figure 3: LINE-1 RNA in P-bodies and stress granules co-localizes with autophagosomes and is likely degraded by autophagy with ORF1p.

From: Autophagy supports genomic stability by degrading retrotransposon RNA

Figure 3

(a) Fluorescent microscopy of GFP-ORF1p and LC3. (b) Relative number of foci of GFP-ORF1p, or number of foci of GFP-ORF1p co-localized with LC3 per cell in randomly selected fields of cells treated with Bafilomycin A1 (20 h, 400 nM, n=30 cells, total number of GFP-ORF1p foci: control=150, BAF=214, total number of GFP-ORF1p co-localized with LC3: control=32, BAF=199). *P=0.007 Total, P=1 × 10−42 Co-localized with LC3, t-test. Li’s correlation coefficient of co-localization (0.328, s.e.m. 0.019). Error bars represent s.e. of the mean. (c) Fluorescent microscopy of endogenous ORF1p and HcRed-LC3. (d) Western blot analysis of ORF1p in autophagosome-enriched fractions. LC3-II is a marker of autophagosomes. (e) Western blot analysis of GFP and alpha-tubulin (control) in cells transfected with GFP-ORF1p and treated with control or Bafilomycin (20 h, 400 nM). (f) Fluorescent microscopy of cells expressing LINE-1-MS2, MS2-GFP and mCherry-TIA-1. (gi) Fluorescent microscopy of endogenous LINE-1 RNA in cells transfected with GFP-DCP1A (g), GFP-GE-1 (h) or GFP-TIA-1 (i). All experiments were performed with a minimum of three biological replicates.

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