Fig. 4

Knockdown of AKAP4 gene reduces cellular motility and induces cellular senescence in CRC cells. a Representative photomicrographs show the reduction in COLO 205 and HCT 116 cells migrating/invading through transwell membrane when transfected with AKAP4 shRNA3 compared to NC shRNA. Histogram depicts significant reduction in number of cells migrating/ invading through insert membrane when transfected with AKAP4 shRNA3 as compared to NC shRNA. b Western blot analysis of the molecules of EMT demonstrates up regulation of epithelial marker E-cadherin, while down regulation of mesenchymal markers like P-cadherin, N-cadherin, SLUG, α-SMA, SNAIL, TWIST, Vimentin and MMP 2, 3, 9. β-actin was used as a loading control .c Senescence assay: representative phase contrast microscopic images showed higher β-galactosidase activity (green staining) in AKAP4 shRNA3 transfected COLO 205 and HCT 116 cells as compared to NC shRNA transfected cells. Histogram depicts the quantitative difference in the β-galactosidase activity in AKAP4 shRNA3 transfected COLO 205 and HCT 116 cells as compared to NC shRNA transfected cells. Western blot analysis reveals the up-regulation of senescence marker, DCR2, in AKAP4 shRNA3 transfected COLO 205 and HCT 116 cells as compared to NC shRNA transfected cells. *P < 0.05, **P < 0.001, ***P < 0.0001