Fig. 8.

D347-2761 disturbed c-Myc stability to regulate downstream gene transcription. A Indicated cells were treated by 10 μM D347-2761 at different time (12 h, 24 h, 48 h and 72 h) and western blot analysis of c-Myc expression. B Indicated cells were treated by 10 μM D347-2761 following 10 μM MG132 treatment for 4 h, 24 h later, western blot assay of c-Myc expression. C RPMI-8226 and NCI-H929 cells were dealt with 10 μM D347-2761 or not for 6 h and then 5 μg/ml CHX were added into the medium for 20 min and 50 min respectively. Western blot anaylsis of c-Myc protein expression. D RPMI-8226 cells were treated by different dose of D347-2761 (2 μM, 5 μM and 10 μM) for 48 h, western blot analysis of p-c-Myc Thr58, p-c-Myc Ser62, p-GSK3-β and GSK3β. β-actin was used to be internal control. E Real time PCR analysis of potential c-Myc target genes mRNA levels in RPMI-8226 cells via D347-2761 treatment. F qChIP was performed to measure the levels of c-Myc at CDK4 promoter in RPMI-8226 cells after D347-2761 treatment. The experiments were repeated at least three times. Error bars: mean ± SD. *P < 0.05, **P < 0.01