Fig 2: JND4135 induces apoptosis and G0/G1 cell cycle phase arrest in BaF3-TRKA-G667C cells. (A) JND4135 induces apoptosis in BaF3-CD74-TRKA-G667C cells. Cells were treated with indicated compounds for 48 h before Annexin V and 7-AAD co-staining. (B) JND4135 induces G0/G1 cell cycle arrest in BaF3-CD74-TRKA-G667C cells. Cells were treated with indicated compounds for 24 h before DNA labeling by propidium iodide. (C) The effect of JND4135 on the expression of p-TRKA, CDK2, CDK4, Cyclin D2, Caspase-9, Caspase-3 and PARP was tested by Western blotting. The cells were treated with JND4135 from 0.4 nM to 10 nM for 48 h. Immunoblot data are representative of three independent experiments.

Fig 3: SOX2 was found to regulate cell cycle and apoptosis of Ewing’s sarcoma cells via the PI3K/Akt signaling pathway. a WB assay showed SOX2 knockdown resulted in decreased expressions of PI3K and Akt and the loss of p-Akt in A673 and RD-ES cells. b Overexpression of Akt was evaluated by WB analysis. c Four groups were designed for Ewing’s sarcoma cells as mock control, SOX2 knockdown, Akt overexpression and the contransfection of siSOX2 and Akt plasmids. Expression levels of SOX2, Akt and p-Akt of the four groups were examined by WB. d Further investigation of the proteins targeted by SOX2 and Akt involved in the Ewing’s sarcoma cells apoptosis and G1/S transition, including Fas, Bad, caspase-9, caspase-3, p21, p27 and cyclin-E, were analyzed by WB in the four designed groups. Representative data from one of three independent experiments are exhibited in (a–d)

Fig 4: Zinc inhibits ZEA-induced cell apoptosis.The effects of ZnSO4, GZn, and ZEA on the ??m (A) and the percentage of cell apoptosis (B). The ??m is expressed as the fluorescence intensity ratio of the red over the green staining. The effects of ZnSO4, GZn, and ZEA on the mRNA expression of Bax (C), Casp3 (D) and Casp9 (E). KK-1 cells were treated as described in the legend for Fig. 1. The values are the means ± SD of three independent experiments. Different characters indicate significant differences between the compared groups (p < 0.05).

Fig 5: EGCG reversed H2O2-induced up-regulation of apoptosis marker proteins. (A) Representative western blot analysis of cleaved-caspase 3, cleaved-caspase 9 in EGCG (1, 5, 10 µmol/L) pretreated HUVECs. Densitometric analysis was used to quantify the level of (B) cleaved-caspase 9 and (C) cleaved-caspase 3 activation. Values were normalized against ß-actin and presented as fold increases compared with those at the basal level (control). Data are shown as the mean ± SEM, n=3. *P<0.05 vs. control group, **P<0.01, ***P<0.001 vs. H2O2 alone group.