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Fig. 1 | Molecular Brain

Fig. 1

From: Development of GABARAP family protein-sensitive LIR-based probes for neuronal autophagy

Fig. 1

Efficient localization of HyD-2xLIRs-GFP to mRFP-LC3/GABARAP-positive autophagosomes. a Schematic model depicting the development of new autophagosome sensors. LIR: LC3-interacting region, HyD: hydrophobic domain. b Confocal images showing the cellular localization of HyD-LIR(X)-GFP or HyD-2xLIR(X)-GFP together with mRFP-LC3A/B/C and GABARAP/−L1-L2 in MEFs incubated with 100-nM rapamycin (Rapa) + 10-mM NH4Cl for 4 h. Scale bar: 10 μm. c The bar graphs illustrate the ratio of autophagosomal/cytosol (A/C) GFP fluorescence intensity in cells expressing HyD-LIR(X)-GFP or HyD-2xLIR(X)-GFP and each mRFP-LC3/GABARAP protein. d The LC3/GABARAP protein binding properties of HyD-2xLIR(Stbd1)-GFP or HyD-2xLIR(ULK2)-GFP assessed using GST pull-down assays. e Confocal images showing cellular localization of HyD-2xLIR(Stbd1)-GFP or HyD-2xLIR(ULK2)-GFP in wild-type (WT), ATG5-knockout (ΔATG5), or ATG7-knockout (ΔATG7) HeLa cells treated with 100-nM rapamycin (Rapa) + NH4Cl. Scale bar: 10 μm. f Confocal images showing cellular localization of HyD-2xLIR(Stbd1)-GFP together with mRFP-LC3B or mRFP-GABARAP-L1 in cultured cortical neurons incubated in the absence or presence of 50 μM chloroquine (CQ) for 1 h. Scale bar: 10 μm. X; Stbd1 or ULK2

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