Figure 2 | Scientific Reports

Figure 2

From: Ubiquitin ligase ITCH recruitment suppresses the aggregation and cellular toxicity of cytoplasmic misfolded proteins

Figure 2

Interaction of ITCH with misfolded proteins.

(A–B) Transient transfection was used to overexpress GFP-wtCAT and GFP-Δ9CAT constructs and the cells were processed for immunofluorescence staining with nuclear staining using DAPI (A). The same sets of cell lysates were probed with an anti-ubiquitin antibody (B). (C) GFP-wtCAT- and GFP-Δ9CAT-transfected cell lysates were pulled down by an anti-ITCH antibody and the blots were developed using an anti-GFP antibody (upper panel) and anti-ITCH antibody (lower panel). (D) The same transfected samples were processed for immunoprecipitation with the anti-GFP antibody and the blots were processed for immunoblotting using anti-ITCH (upper panel) and anti-GFP antibodies (lower panel). (E–F) The cells were transfected as described in A and 48 h after transfection, cell lysates were prepared and processed for immunoprecipitation (IP) using beads only (control); immunoprecipitates were then probed with anti-GFP (E) and anti-ITCH antibodies (F).

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