Fig. 2

Ablation of AKAP4 protein alters malignant properties of CRC cells. a Western blot show knockdown efficiency of shRNA targets against AKAP4 protein in COLO 205 and HCT 116 cells. b Histograms depict the effect of AKAP4 shRNA3 on cellular proliferation and cell viability of COLO 205 and HCT 116 cells as compared to NC shRNA at 24 h, 48 h and 72 h. c Histogram depicts the difference between the number of colonies being formed in AKAP4 shRNA3 transfected COLO 205 and HCT 116 cells as compared to NC shRNA transfected COLO 205 and HCT 116 cells. d The blots show changes in the expression of various molecules that are involved at different phases of the cell cycle in COLO 205 and HCT 116 cells when transfected with AKAP4 shRNA3 as compared to NC shRNA. Upregulation of p16, p21, Rb protein while down regulation of Cyclins namely Cyclin B1, D, E and cyclin dependent kinases (CDK’s), CDK 1, 2, 4, 6 and phosphorylated Rb was observed. Proliferation marker PCNA was also down regulated. β-actin was used as a loading control. e Flow cytometric analysis reveals DNA fragmentation as assessed by TUNEL assay. Surface expression of phosphotidyl serine was assessed by AnnexinV-PerCP-Cy5-5-A staining. The blue peak shows the cells transfected with NC shRNA while the red peak shows the cells transfected with AKAP4 shRNA3. *P < 0.05, **P < 0.001, ***P < 0.0001