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. 2018 May 7;217(5):1757–1776. doi: 10.1083/jcb.201708168

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© 2018 Sha et al.

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Figure 8. — p62 knockdown in neuroblastoma cells did not reduce protein degradation or increase levels of Ub conjugates but impaired the buildup of polyubiquitinated and polysumoylated proteins in inclusions. (A) p62 knockdown (sh-p62) did not affect the ability of SH-SY5Y cells to degrade long-lived proteins when treated or not with 1 µM BTZ to completely inhibit the proteasome. n = 6. (B) In M17 cells treated with BTZ, stable knockdown of p62 but not Nbr1 or GABARAPL1 reduced the buildup of Ub or SUMO2/3 conjugates in the pellet that contains inclusions. In the supernatant, there are no SUMO2/3-conjugated proteins, and the bands recognized by the SUMO2/3 antibody (asterisk) are not specific. (C and D) In SH-SY5Y cells treated for 16 h with 100 nM BTZ, knockdown of p62 reduced Ub (C) and SUMO2/3 (D) conjugates in the pellet. (C) Knockdown of Nbr1 and GABARAPL1 also reduced Ub conjugates in the pellet. Molecular masses are given in kilodaltons. Error bars indicate SD.