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Figure 1 | Journal of Molecular Signaling

Figure 1

From: Defining regulatory and phosphoinositide-binding sites in the human WIPI-1 β-propeller responsible for autophagosomal membrane localization downstream of mTORC1 inhibition

Figure 1

Generation and transient expression of GFP-WIPI-1 mutants in human U2OS cells. (A) Representation of the human WIPI-1 protein harbouring 7 WD repeats (coloured boxes 1–7) and evolutionarily conserved amino acids (homologous amino acids in red, invariant amino acids in purple, not conserved residue in black). (B) Transient over-expression and anti-GFP ECL analysis of GFP-tagged wild-type WIPI-1 (labelled WIPI-1) or generated mutant WIPI-1 variants (labelled N23A – G336) along with the GFP control and untransfected U2OS cells. The RR mutant carries alanine substitutions for both R226 and R227. Supplementary material provides an amino acid sequence alignment of wild-type and mutant WIPI-1 variants deduced from automated DNA sequencing upon site-directed mutagenesis (Additional file 1: Figure S1).

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