Skip to main content
. 2019 Mar 11;47(9):4476–4494. doi: 10.1093/nar/gkz163

Figure 3.

Figure 3.

GSK3β is required for IRF1 transcriptional activity. (A) Reporter assays in Cos7 cells transfected with IRF1 and TRAIL promoter reporter for 48 h. Cells were treated with NaCl (to control for osmolality) or LiCl for 24 h prior to lysis. Data is expressed as fold luciferase induction by IRF1 over empty vector (pCDNA3.1). All reporter assay data is from three independent experiments assayed in triplicate. Error bars denote SEM and * denotes statistical significance (P < 0.05) as determined by Students t-test between NaCl and LiCl treatments. Panel below shows IRF1 expression. (B) As for (A) but treatment with vehicle (DMSO), GSK3 Inhibitor BIO or the inactive analog Methyl-BIO (1, 2.5 and 3.75 μM/1 h). (C) Reporter assays using Cos7 cells transfected with TRAIL reporter construct, pcDNA3 (vector), or IRF1 and increasing concentrations of GSK3β-HA WT or GSK3β-HA K85A mutant. (D) Reporter assays in MRC5 cells transfected with control or GSK3β siRNAs (10 nM/16 h) followed by transfection with TRAIL promoter reporter and IRF1 for 24 h. (E) Reporter assay in Cos7 cells transfected with the TRAIL or 4XISRE-Luc reporters in conjunction with IRF1 WT, T181A, S185A and T181A/S185A constructs. Statistical difference is between WT and mutant IRF1. (F) As for (E) but with T181D and S185E mutants in Cos7 and MRC5 cells.