Fig. 2.

IR5 contains a conserved PU.1 consensus that is required for transcription from the LIMD1 promoter. (A) LIMD1 promoter sequences from different LIMD1 expressing species was extracted from the Ensembl genome browser and aligned using ClustalW. Perfect homology is observed at the IR5 and within the IR7 consensus. Further scrutinisation of IR5 using MatInspector identified a putative PU.1 binding consensus sequence. (B) Point mutagenesis of the PU.1 binding consensus sequence compared to the WT consensus are shown (CC-TT and C-T). In addition we performed reverse mutagenesis to restore the PU.1 consensus (TT-CC) and reverse single conserved mutation outside of this consensus (T-C). All mutated promoter reporters together with WT and IΔ5 LIMD1 promoters were analysed in reporter assays as in Fig. 1B. Bold – PU.1 consensus; italics and underlined – mutated bases.