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. 2020 May 31;12(6):1427. doi: 10.3390/cancers12061427

Figure 4.

Figure 4

RL2 interacts with mitochondrial membrane protein TOM70. (A) Workflow for RL2-based mass spectrometry analysis. MDA-MB-231 cell lysates were incubated with RL2-coupled Protein A Sepharose beads (RL2-pulldown) or Protein A Sepharose beads (control). Precipitated proteins were identified by nanoLC-tandem mass spectrometry. (B) Mass spectrometry identified unique peptides for each protein. The median of the unique peptides from three independent experiments is shown for each protein and compared for RL2- and control pulldown. The scoring indicates the increase in unique peptides identified. (C) RL2-pulldown precipitates were analyzed by Western Blot and probed for indicated proteins. (D) MDA-MB-231 and (E) MCF-7 cells were treated with 200 µg/mL RL2 as indicated. TOM70 immunoprecipitation (TOM70 IP) was performed with TOMM70A antibody or bead-only control and probed for indicated proteins by Western Blot. One representative experiment out of three is shown for all Western Blots and immunoprecipitations are shown in Figure S3.