Modulating effects of acyl-CoA synthetase 5-derived mitochondrial Wnt2B palmitoylation on intestinal Wnt activity

Figure 2 Wnt activation dependent on acyl-CoA synthetase 5 expression. A: Luciferase reporter assay in HEK293 and CaCo₂ cells stably or transiently transfected with acyl-CoA synthetase 5 (ACSL5) and transiently transfected with pTOPflash or pFOPflash. After activation with Wnt3A, Tcf reporter-derived luciferase activity was detected luminometric; B: Luciferase reporter assay after knockdown of ACSL5 by siRNA in HEK293 and CaCo₂ cells; C: Luciferase reporter assay after knockdown of Wnt2B by siRNA in HEK293 cells; D: Palmitoylation of Wnt2B dependent on ACSL5 expression. Isolated mitochondria from HEK293 cells, transiently transfected and immunoprecipitated with Wnt2B, with or without treatment with palmostatin B, Hydroxylamine (hydrochloride), and BMCC. As a reference, Wnt2B protein expression was determined; E: Isolated mitochondria from CaCo₂ cells, transiently transfected with Wnt2B and ACSL5, immunoprecipitated with Wnt2B, after treatment with palmostatin B, hydroxylamine (hydrochloride), and BMCC. ^aP < 0.05 vs control.