Figure 1.
Apigenin inhibits the viability of ANA-1 cells.
The ANA-1 cells were exposed to either vehicle (2 mM Na2CO3 in medium) or apigenin and were incubated as described above. Cell viability was analyzed by the MTT assay. All data are expressed as the percentage of change in comparison with the control group, which were assigned 100% viability. The values are given as the mean ± SD. *p<0.05, **, p<0.01, versus the control.
Figure 2.
Apigenin induces the apoptosis of ANA-1 cells.
A. The quantification of flow cytometry analyses. The flow cytometry data showed one representative sub-G1 result. B. The apoptotic rates are given as the percentage of cells with sub-G1 DNA content. The values are given as the mean ± SD. *, p<0.05, versus the control. C. ANA-1 macrophages were exposed to different concentrations of apigenin for 24 h and 48 h. The flow cytometry data showed one representative dual staining result. D. Early- (%) and late-stage (%) apoptotic cell death were detected by staining cells with Annexin V-FITC and PI and analyzing by flow cytometry. The values are given as the mean ± SD. *, p<0.05, **, p<0.01, versus the control. E. the ANA-1 cells were exposed to apigenin for 48 h. The images of TUNEL positive cells were captured by flow cytometry and a confocal microscopy (×200).
Figure 3.
Apigenin induced apoptosis in ANA-1 cells.
A typical DNA ladder in ANA-1 cells was observed after being treated with 50 μM apigenin for 48 h. Lanes: 1) control; 2) 12.5 μM apigenin; 3) 25 μM apigenin; 4) 50 μM apigenin.
Figure 4.
The effects of apigenin on apoptotic and anti-apoptotic proteins in ANA-1 cells.
A. Representative western blot results showed Bcl-2, Bax, caspase-3 and caspase-8 expression in the ANA-1 cells after incubation with apigenin for 48 h. B. The relative expression of proteins compared with the control, *, p<0.05, **, p<0.01, versus the control.
Figure 5.
Effect of apigenin on intracellular ROS levels in the ANA-1 cells.
The ANA-1 cells were treated with apigenin for 24 h. The ROS levels in the ANA-1 cells. A. The ROS levels are given as the MFI of DCF of cells. The values are given as the mean ± SD. *, p<0.05, **, p<0.01, versus the control. B. Representative FACS results showed apigenin increased the intercellular ROS levels in the ANA-1 cells after 24 h.
Figure 6.
The effects of apigenin on the MAPK pathway in ANA-1 cells.
A–C. Representative western blot results showed phospho-ERK1/2/ERK1/2, phospho-JNK/JNK and phospho-p38/p38 expression in the ANA-1 cells after incubation with apigenin for 48 h. D. The relative expression of proteins compared with the control, *, p<0.05, **, p<0.01, versus the control.