FIGURE 3.

DYRK1A elevated NFATC1 protein expression and transcriptional activity. A, DYRK1A increased protein expression of NFATC1 in T98G cells. T98G cells were transfected with pCMV‐DYRK1A. NFATC1 and DYRK1A were detected by NFATC1 monoclonal antibody and DYRK1A polyclonal antibody. ACTB was used as the loading control. B, Inhibition of DYRK1A decreased protein expression of NFATC1 in T98G cells. T98G cells were transfected with pshDYRK1A. NFATC1 and DYRK1A were detected using NFATC1 monoclonal antibody and DYRK1A polyclonal antibody, respectively. ACTB was used as the loading control. C, NFATC1 target genes’ transcripts were increased by DYRK1A overexpression. T98G cells were cotransfected with p‐NFATC1mycflag and pCMV‐DYRK1A. qRT‐PCR was used to detect the mRNA expression of NFATC1 targets IL2 and TNFα. D, The mRNA levels of NFATC1 target genes IL2 and TNFα were decreased by DYRK1A knockdown. E, NFATC1 transcriptional activity was increased by DYRK1A. T98G cells were co‐transfected with NFATC1 activity reporter pNFATluc, p‐NFATC1mycflag, and pCMV‐DYRK1A. Dual luciferase assay was performed 48 h after transfection. F, Inhibition of DYRK1A decreased NFATC1 transcriptional activity. G, Cytokine array showed differential cytokine expression profiles regulated by DYRK1A in T98G cells. T98G cells were transfected with p‐NFATc1mycflag and pCMV‐DYRK1A. A RayBiotech human cytokine antibody array was performed to indicate transcriptional activity of NFATC1 48 h after transfection. Values represent means ±SD, n = 3. p < 0.05 by student t‐test