Fig. 6: 14-3-3ζ involved in the regulation of LCK on the multimerization of TRPM8. | Cell Death & Disease

Fig. 6: 14-3-3ζ involved in the regulation of LCK on the multimerization of TRPM8.

From: The LCK-14-3-3ζ-TRPM8 axis regulates TRPM8 function/assembly and promotes pancreatic cancer malignancy

Fig. 6: 14-3-3ζ involved in the regulation of LCK on the multimerization of TRPM8.

A, B PANC-1 cells were co-transfected Flag-TRPM8 with HA-LCK or control vector and then harvested for IP with an anti-Flag antibody and WB assay with the indicated antibodies to determine the binding of TRPM8 and 14-3-3ζ. C, D Similar experiments in (A) and (B) but cells co-expressing Flag-TRPM8 along with siLCK. E, F Expression constructs for Flag-WT-TRPM8 or Flag-TRPM8-Y1022F were co-transfected with or without HA-LCK into HEK293T cells to determine the binding of TRPM8 and 14-3-3ζ. G, H PANC-1 cells were co-transfected with Flag-TRPM8, HA-LCK, si14-3-3ζ (#1, #2 or #3) or their combination, before harvest for treatment with 1 μM DSS for 30 min, and subjected to WB assay to detect the level of TRPM8 multimerization. I, J AsPC-1 cells were co-transfected with Flag-TRPM8 and GFP-TRPM8, HA-LCK, si14-3-3ζ or their combination, and then harvested for IP with an anti-Flag antibody and subjected to determine the binding of intermolecular TRPM8. ***P < 0.001, NS not significant. Data are presented as mean ± SEM. All studies were repeated at least three times.

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