Fig 1: CKAP2L is overexpressed in ESCC tissues and cell lines. (a) Volcano plot for differentially expressed genes in ESCC on the basis of GSE17351. The fold change (FC) of genes was assessed by log transformation. |logFC| >2.5 and adjusted P < 0.05 were defined as the screened threshold. (b) The data were from TCGA & GTEx database and analyzed by the GEPIA website, the expression of CKAP2L was upregulated in tumor tissues compared with normal tissues (P < 0.05). (c) Expression levels of CKAP2L in clinical samples were detected by RT-qPCR. *P < 0.05 by student's T-test. (d) Expression levels of CKAP2L in ESCC cell lines were detected by RT-qPCR. *P < 0.05 by student's t-test versus Het-1A. (e) Western blot analyses the protein expression of CKAP2L in ESCC cells compared with the normal cells Het-1A. GAPDH was used as a loading control in the western blot. The data were expressed as mean ± SD (n = 3), *P < 0.05.
Fig 2: CKAP2L expression positively correlates with the malignant proliferation of ESCC cells in vivo. (a) Effect of subcutaneous injection of KYSE150 and Eca109 cells transfected with shCKAP2L or CKAP2L-OE on the tumor growth (n = 4). (b) The mean of tumor diameters for every week is presented *P < 0.05 by ANOVA. (c) Quantitative analysis of tumor weight *P < 0.05 by student's t-test. (d) Immunostaining analysis was performed to measure the positive rate of Ki67 in xenograft tissues. (e) TUNEL assay was performed to determine the rate of cell apoptosis. *P < 0.05 by student's t-test. The data were expressed as mean ± SD (n = 3).
Fig 3: Expression of CKAP2L inhibits the sensitivity of ESCC cells to flavopiridol. (a), (b) IC50 of flavopiridol was determined by CCK-8 assay upon flavopiridol treatment for 24 h in shCKAP2L and CKAP2L-OE transfected KYSE150 and Eca109 cells *P < 0.05 by ANOVA and student's t-test. (c), (d) CCK-8 assay was used to detect the proliferation activity of shCKAP2L and CKAP2L-OE cells after treatment with flavopiridol. *P < 0.05 by ANOVA. (e) Cell apoptosis of shCKAP2L and CKAP2L-OE cells was detected after flavopiridol (5 µM) treatment for 48 h through FITC-Annexin V/PI assays and flow cytometry. The data were expressed as mean ± SD (n = 3).
Fig 4: CKAP2L facilitates migration and invasion of ESCC cells in vitro. (a), (b) Wound healing assays were performed to detect the effect of shCKAP2L and CKAP2L-OE on cell-migration ability. (c), (d) The effects of the CKAP2L expression on the invasiveness of KYSE150 and Eca109 cells were investigated in transwell assays. *P < 0.05 by student's t-test. The data were expressed as mean ± SD (n = 3).
Fig 5: CKAP2L promotes ESCC cell proliferation in vitro. (a) The expression of CKAP2L was knockdown by siRNAs in KYSE150 cells. *P < 0.05 by student's t-test versus si-Ctrl. (b) Western blotting was used to measure the expression of CKAP2L after knockdown or overexpression by shCKAP2L and CKAP2L-OE. *P < 0.05 by student's t-test versus shCtrl. (c), (d) CCK-8 and colony formation assays were performed to determine the proliferation ability of ESCC cells transfected with shCKAP2L and CKAP2L-OE. And, the colony number was counted. *P < 0.05 by student's t-test. (e) EdU analysis displayed that cell proliferation was increase by CKAP2L in ESCC cells. The data were expressed as mean ± SD (n = 3), *P < 0.05.
Supplier Page from Abcam for Anti-CKAP2L antibody