Fig. 2. HOXA-AS3 knockdown reduces cell proliferation and regulates the cell cycle.

a The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell viability in A549 cells. b 5-bromodeoxyuridine incorporation showed the synthesis of DNA. c Western blot analyses of the protein expression levels of proliferating cell nuclear antigen. d Immunofluorescence for ki67 in A549 cells, after staining the nuclei with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 25 μm. e Left, fluorescence-activated cell sorting (FACS) analyses detected the number of cells in each phase of the cell cycle. Right, the percentage of cells in the S and G2/M phases in A549 cells. f Western blot analyses of the protein expression levels of cyclin (a), cyclin (d), and cyclin (e). g Western blot analyses of the protein expression levels of CDK1, 2, and 4. NC, negative control; siRNA/HOXA-AS3, small interfering RNA for HOXA-AS3. ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001. All values are expressed as the mean ± SEM