Figure 5: GSK3 inhibition leads to NF-kB activation and its binding to the TNF promoter in human NK cells.

(a) Human NK cells were treated with the GSK3 inhibitor SB for the indicated times, cytoplasmic and nuclear lysates were prepared, and analysed by Western blotting using indicated antibodies. Actin and hnRNPA1 were used as internal loading controls for cytoplasmic and nuclear extracts. Tubulin was used as a control to examine the purity of nuclear extracts. Experimental control in which NK cells were treated with DMSO, the vehicle used to dissolve SB is shown in Supplementary Fig. 6. (b) Expanded NK cells were treated with SB for the indicated times. Nuclear extracts were prepared and analysed by Oligonucleotide pull-down assay to examine NF-kB binding to the TNF promoter. Data presented are representative of two independent studies. Experiment is performed three times.