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. 2015 Mar 10;6(10):7992–8006. doi: 10.18632/oncotarget.3505

Fig.7. The activation of Notch1 signaling is involved in tetrandrine-induced NB4 cell autophagy and differentiation.

Fig.7

(A) RT-PCR analysis of the relative mRNA expression levels of HES1, HES5 and Notch1. NB4 cells were exposed to 2μM tetrandrine for 24 hours. Oxidant dimethylsulfoxide (DMSO) was used as a negative control (Con). Error bars represent the mean ±SD. **p <0.01. (B) Western blot analysis of NICD and HES1 protein levels after tetrandrine (Tet) treatment at the indicated doses and time intervals. (C) Western blot analysis of HES1 and LC3 levels. NB4 cells were treated with DMSO (Con), 2 μM tetrandrine (Tet), 2 mM DAPT, and 2 μM tetrandrine (Tet) after a 1-hour pretreatment with 2 mM DAPT (Tet+ DAPT) for 24 hours. (D) Acridine orange staining assay analysis of autophagy. NB4 cells treated with 2 μM tetrandrine (Tet) after a 1-hour pretreatment with 2 mM DAPT. Error bars represent the mean ±SD. **p <0.01. (E) NB4 cells were 1-hour pretreated with DAPT and incubated with 2 μM tetrandrine (Tet) for 4 days prior to CD14 detection by flow cytometry.