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. 2016 Dec 27;36(4):549–564. doi: 10.15252/embj.201695063

Figure 5. Performance of ATG8 sensors can be increased by introduction of the oligomerization domain PB1.

Figure 5

  1. LC3C‐specific sensor mCh‐AS3_67 is recruited to Salmonella (SFH2) escaping SCVs. Salmonella SFH2 strain is carrying a GFP‐reporter gene activated by availability of glucose‐6‐phosphate in the host's cytosol.
  2. Salmonella with prolonged cytosolic residence and strong activation of the GFP‐reporter gene (green arrow) are not recognized by the LC3C‐specific sensor mCh‐AS3_67.
  3. A subpopulation of hyperproliferative Salmonella in the cytosol is coated with endogenous LC3C. In contrast, endogenous LC3B is enriched in spots in close proximity to the cytosolic bacteria.
  4. Sensitivity of LC3C‐specific sensor mCh‐AS3_67 is increased upon fusion of membrane‐tethering FYVE domain or PB1 oligomerization domain.
  5. Introduction of the PB1 domain into mCh‐AS3_p62 increases its performance in recognizing endogenous LC3B.
  6. High‐performance sensor mCh‐PB1‐AS3_67 was transiently expressed in Salmonella‐infected HeLa cells to monitor xenophagy events.
Data information: All experiments were performed in HeLa cells, except for (E), which was performed in U2OS cells. Scale bars: 10 μm.