Fig. 1.

MLN4924 induced ATF3 accumulation at the transcriptional level. a EC1 and Kyse450 cells were treated with MLN4924 for 24 h and cell lysates were assessed by IB with specific antibody against ATF3. b EC1 and Kyse450 cells were treated with MLN4924 for 0 h, 12 h, 24 h or 48 h with 0.6 μmol/L MLN4924 and cell lysates were assessed by IB with specific antibody against ATF3. c-f MLN4924 did not enhance the protein stability of ATF3. EC1 and Kyse450 cells were pretreated with 0.6 μmol/L MLN4924 for 12 h to increase the basal protein level of ATF3. Cells were then washed with PBS to remove the residual of MLN4924 and divided into two groups, which were further treated with 50 μg/mL of cycloheximide in the presence or absence of MLN4924 (0.6 μmol/L) for indicated period of time and then collected for IB analysis. The protein level was quantified by densitometric analysis using Image J software. g-h MLN4924 increased ATF3 at the transcriptional level. EC1 and Kyse450 cells were treated with MLN4924 for 0 h, 12 h, 24 h or 48 h with 0.6 μmol/L MLN4924 and RNA were extracted and analyzed by qPCR (normalized to GAPDH)