Fig 1: Exploration of downstream molecular mechanism of ALPK2 in OC cells. a Human apoptosis antibody array analysis was performed in HO-8910 cells with or without ALPK2 knockdown. b Densitometry analysis was performed and the gray values of differentially expressed proteins were shown. c The expression of epithelial-mesenchymal transition (EMT) proteins were observed by Western blot in HO-8910 and OVCAR-3 cells. d The expression of target proteins pathways were observed by Western blot in HO-8910 cells. The data were expressed as mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001
Fig 2: Knockdown of ALPK2 inhibits tumor growth in mice xenograft models. a The volume of tumors in shCtrl group and shALPK2 group was measured post-injection. b The average weight of tumors in shCtrl group and shALPK2 group. c The photos of tumors in shCtrl group and shALPK2 group. d The total bioluminescent intensity of tumors in shCtrl group and shALPK2 group. e The bioluminescence imaging of tumors in shCtrl group and shALPK2 group. f Images of the tumor tissues of the shCtrl group and shALPK2 group with Ki-67 staining. The data were expressed as mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001
Fig 3: ALPK2 is highly expressed in OC tissues and the construction of ALPK2 knockdown cell model. a The expression of ALPK2 in the normal and tumor samples detected by IHC. b Transfection efficiency for HO-8910 and OVCAR-3 cells was evaluated by expression of green fluorescent protein 72 h post-infection. c, d The specificity and validity of the lentivirus-mediated shRNA knockdown of ALPK2 expression was verified by qRT-PCR (c) and Western blot (d). The data were presented as the mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001
Fig 4: Depletion of ALPK2 attenuated cell proliferation, migration, and colony formation of esophageal cancer cells. (a) Evaluation of the rate of cell proliferation in esophageal cancer cell lines following infection was determined by the MTT assay. (b) The cell migration rate was detected in esophageal cancer cell lines after being transfected via transwell assay. Magnification times: 200x. (c) The cell migration rate was detected in esophageal cancer cell lines after infection by wound-healing assay. (d) The ability of colony formation was evaluated in esophageal cancer cell lines following infection. The findings were represented as mean ± SD. ***P < 0.001.
Fig 5: Knockdown of ALPK2 inhibits cell proliferation, cell cycle and migration, promotes apoptosis in OC cells. a Cell proliferation of HO-8910 and OVCAR-3 cells with or without knockdown of ALPK2 was evaluated by MTT assay. b The HO-8910 and OVCAR-3 cells in the G1, S and G2 phases of the ALPK2 gene knockdown group and the control group were detected by flow cytometry. c Flow cytometry analysis based on Annexin V-APC staining was utilized to detect the percentage of early apoptotic cell for HO-8910 and OVCAR-3 cells. The X axis indicated the cell apoptosis while the Y axis indicated the green fluorescence detected from the GFP tagged on lentivirus (shALPK2 or shCtrl). d Cell migration of HO-8910 and OVCAR-3 cells with or without knockdown of ALPK2 was evaluated by wound-healing assay. The data were expressed as mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001
Supplier Page from Abcam for Anti-ALPK2 antibody