Fig. 6: ERK5i induce expression from the KLF2 promoter and is dependent on ERK5 TAD.
From: Paradoxical activation of the protein kinase-transcription factor ERK5 by ERK5 kinase inhibitors
a HEK293 cells were transfected with KLF2:LUC and CMV:Renilla together with either FLAG or FLAG-MEF2D and either HA or HA-ERK5 (full length), and either EGFP (control) or EGFP-MEK5D as indicated. Twenty-four hours post transfection, cells were lysed and firefly luciferase activity was measured and normalised to Renilla. The results are presented as the mean of three independent experiments ± SEM. Source data are provided as a Source Data file. b HEK293 cells were transfected with KLF2:LUC and CMV:Renilla together with FLAG and HA, with either EGFP or EGFP-MEK5D (to activate endogenous ERK5). Four hours post transfection, cells transfected with KLF2:LUC and CMV:Renilla together with FLAG, HA and EGFP were treated with either compound 26, AX15836 or DMSO (control) at the concentrations indicated. Cells transfected with KLF2:LUC and CMV:Renilla together with FLAG, HA and EGFP-MEK5D were treated with DMSO. Twenty-four hours post transfection, cells were lysed and firefly luciferase activity was measured and normalised to Renilla. The results are presented as % KLF2 promoter activity where MEK5D-driven (through endogenous ERK5) KLF2 promoter activity is 100% (mean of three independent experiments ± SEM). Source data are provided as a Source Data file. c HEK293 cells were transfected with KLF2:LUC and CMV:Renilla, together with FLAG-MEF2D and either HA-ERK5 (full length) or HA-ERK5 ΔTAD, and either EGFP (control) or EGFP-MEK5D. Four hours post transfection cells were treated with either 300 nM AX15836 or DMSO (control). Twenty-four hours post transfection cells were lysed and processed as in b.
