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. 2019 Feb 25;38(6):e100024. doi: 10.15252/embj.2018100024

Figure EV1. Generation and analysis of NEDP1 knockout U2OS cells.

Figure EV1

  1. Western blot analysis of whole‐cell lysates from U2OS WT and NEDP1 KO cells reveals a loss of free NEDD8 and an accumulation of NEDD8 reactive species in the NEDP1 KO lysate.
  2. Neddylated species are not dependent on the ubiquitin E1. Western blot analysis of whole‐cell lysate from WT and NEDP1 KO HEK 293 cells treated MLN7243 (0.5 μM) for 8 h. The presence of ubiquitylated species is decreased after treatment, but the amount of neddylated species are not significantly reduced.
  3. Neddylated species are not dependent on the NEDD8 co‐E3s DCNL1‐5. Western blot analysis of whole‐cell lysates from WT and NEDP1 KO HEK 293 cells untreated (NT) or treated with the indicated siRNA for 48 h.