Figure 3.

miR-4634 and RAD001 induce cell apoptosis in vitro. (A) A549 cells were transfected with miR-4634 mimics or NC mimics and treated with RAD001 for 48 h. Apoptotic cells were analyzed by flow cytometry using Annexin V/ PI staining. The flow diagrams were performed using FlowJo V10.0.6 (https://www.flowjo.com/solutions/flowjo/downloads/). (B) Each column indicates the rate of early and late apoptosis of cells, *P < 0.05, **P < 0.01. (C, D) Western blotting was used to detect the apoptosis related proteins and GAPDH was used as a loading control. The western blot analysis were performed using Image Lab V3.0 (https://en.freedownloadmanager.org/Windows-PC/Image-Lab.html). In order to save antibodies and incubate multiple antibodies at once to improve efficiency, we cut the PVDF membrane. The images of western blotting are cropped. The full-length blots/gels are presented in Supplementary Fig.S1-S11.