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. 2022 Sep 20;41(21):e110372. doi: 10.15252/embj.2021110372

Figure EV4. TSG101 interacts with and enzymatically activates PARP1.

Figure EV4

  • A
    Final protein–protein interaction probability scores for the indicated interactions were obtained from the PrePPI database (Zhang et al, 2013).
  • B
    Validation of PARP1 knockdown in A. Different PARP1 siRNAs were used in each independent PLA experiment summarized in Fig 4B. The representative image in Fig 4A was obtained using siPARP1(3).
  • C
    U2‐OS cells were transfected with the full‐length mCherry‐tagged TSG101 plasmid and DNA damage was induced by irradiation (20 Gy, 45 min before analysis), as indicated. Immunoprecipitation of mCherry was performed using whole‐cell extracts. The result is representative of three biologically independent experiments.
  • D
    TSG101 deletion constructs CC+SB and ΔCC were expressed in U2‐OS cells. Untreated or irradiated cells were harvested (90 min after 20 Gy exposure) and analyzed by Western blotting for PAR levels using the Pan ADPr reagent. Vinculin was detected as a loading control.