Fig. 8

A luciferase M4-reporter assay system to study the functional role of Ikaros for M4-harboring genes. a Scheme illustrating the modular M4-reporter gene. The M4 motif is composed of a THAP11 binding site (ACTAYR, purple) and an Ikaros binding site (TCCCRR), yellow) which overlaps with the binding site of the transcriptional activator STAF/ZNF143 (factor-X in green). b M4-reporter gene activity (indicated as relative luciferase units) of M4-WT compared to mutated M4-variant promoters in HEK 239 and HEK293-Ikaros cells, which are stably expressing heterologous Ikaros. Transcriptional activity of corresponding M4-promoters in the absence (black bars) or presence (white bars) of Ikaros. Immunoblot analysis measuring Ikaros protein expression levels and input loading control (tubulin alpha) c Nucleotide sequence presentation of the M4-modules and its variants used for M4-reporter gene assays. d Transient luciferase reporter assay employing M4-reporter Q and R (details: panel c). Ikaros is repressing M4-reporter Q in a dose-dependent manner