GFP-polyQ74 (#40262), GFP-p62 (#38277), GFP-NRF2 (#21549), GFP-KEAP1 (#28025), Flag-KEAP1 (#28023), Myc-NRF2 (#21555), Flag-NRF2 (#36971), HA-K48 (#17605), HA-K63 (#17606), Flag-ULK1 (#27636) were procured from Addgene. GFP-TRIM16 and Flag-TRIM16 were cloned as described previously (Chauhan et al, 2016 (link)). Myc-TRIM16, Myc-TRIM16 deletion constructs, and His-K63-UB were generated using Gateway cloning strategy as per standard protocol (Invitrogen).
The siRNA for NRF2 and p62 were purchased from Sigma, and TRIM16, Ubb, Ube2n siRNA were from Dharmacon. For overexpression experiments, HEK293T cells were transfected using calcium phosphate method as per the manufacturer’s instructions (Profection, Promega). Other cells are transfected using Effectene (Qiagen) or Viafect (Promega) or Interference (Polyplus) as per the manufacturer’s instruction.
The siRNA for NRF2 and p62 were purchased from Sigma, and TRIM16, Ubb, Ube2n siRNA were from Dharmacon. For overexpression experiments, HEK293T cells were transfected using calcium phosphate method as per the manufacturer’s instructions (Profection, Promega). Other cells are transfected using Effectene (Qiagen) or Viafect (Promega) or Interference (Polyplus) as per the manufacturer’s instruction.
