Fig. 5

TC2N interacts with and blocks dual specificity protein phosphatase 3 to enhance CSC-like properties of lung cancer. A Z-dock mode of TC2N identified an interaction surface on DUSP3. The TC2N was represented in light blue; DUSP3 in dark blue. B The extracts from lung cancer cells were subjected to IP with anti-Flag antibody and further analyzed by WB with indicated antibodies. Normal IgG was used as a negative control. Whole-cell lysates were used as a positive control (Input). C The cell extracts from lung cancer cells were subjected to IP with anti-DUSP3 antibody and further analyzed by WB with indicated antibodies. Normal IgG was used as a negative control. Whole-cell lysates were used as a positive control (Input). D The extracts from lung tumor of TC2N engineered mice were subjected to IP with anti-DUSP3 antibody and further analyzed by WB with indicated antibodies. Normal IgG was used as a negative control. Whole-cell lysates were used as a positive control (Input). E The extracts from lung cancer cells stably expressing Flag-NC or Flag-TC2N (full-length and truncation) were subjected to IP with anti-DUSP3 antibody and further analyzed by WB with Flag antibodies. Normal IgG was used as a negative control. Whole-cell lysates were used as a positive control (Input). F In vitro phosphatase assay was performed to detect DUSP3 phosphatase activity using p-nitrophenyl phosphate (pNPP). values are shown as Mean ± SEM. (n = 3). Student’s t-tests, ns, no significance; ***P < 0.001