Fig. 2: Physically interaction of LEF1 with the KDM4A complex. | Cell Death & Disease

Fig. 2: Physically interaction of LEF1 with the KDM4A complex.

From: Carcinogenesis promotion in oral squamous cell carcinoma: KDM4A complex-mediated gene transcriptional suppression by LEF1

Fig. 2

A Normally cultured CAL-27 and SCC-9 cells were fixed and analysed by indirect immunofluorescence using antibodies specific to human LEF1, KDM4A, and N-CoR. Nuclear DNA was stained by 4’,6-diamidino-2-phenylindole (DAPI). Scale bar: 10 μm. B Association of LEF1 with KDM4A complex in CAL-27 and SCC-9. Whole-cell lysates were prepared, co-IP was performed using antibodies against LEF1, KDM4A, or N-CoR, and captured samples were immunoblotted with antibodies against the indicated proteins. IgG served as the negative control. C Association of LEF1 with KDM4A in HEK293T. Co-IP of exogenous EGFP-tagged LEF1 and FLAG-tagged KDM4A in HEK293T was detected. IgG served as the negative control. D GST pull-down experiments performed using bacterially expressed GST-fusion proteins and in vitro-transcribed/translated proteins. E Identification of LEF1 domains required for interaction with KDM4A. The Flag-tagged LEF1 or Flag-tagged truncation constructs of LEF1 were overexpressed in HEK293T cells with EGFP-tagged KDM4A. Whole-cell lysates were prepared, Flag and EGFP expression were detected, and co-IP was performed using antibodies against Flag; then, captured samples were immunoblotted with antibodies against EGFP. Cells with overexpression of pCMV6 plasmid and EGFP-tagged LEF1 served as the negative control. F Identification of KDM4A domains required for interaction with LEF1. The Flag-tagged KDM4A or Flag-tagged truncation constructs of KDM4A were overexpressed in HEK293T cells with EGFP-tagged LEF1. Co-IP was performed as indicated in E.

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