Figure 8.

CR8 triggers transient increase in free Noxa level. SH-SY5Y cells were exposed to 5 µM CR8 at time 0, and cell samples were collected at various times thereafter. Proteins were extracted and resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting with antibodies directed against Mcl-1 (A, total Mcl-1) or Noxa (B, total Noxa). Mcl-1 was also immunoprecipitated with anti-Mcl-1 antibodies, and Mcl-1-bound Noxa was detected by Western blotting (C). The supernatant of the immunoprecipitation reaction was used to detect free Noxa (D).