GST alone, as a negative control, failed to induce the mobility shift (Fig. 3f and Additional file 2: Figure S2b).
To test whether LhrA binds to this region in the chiA mRNA, we performed gel mobility shift assays (Figure 4B).
The fact that Aurora B inhibitors reduce the mitotic phosphorylation of CHMP4C as measured by mobility shifts (PMID 22422861; Figure 4F) is now better explained by the new data indicating that ULK3 is a downstream effector of Aurora B, thus suggesting that Aurora B inhibition is likely to have a knock-on effect on ULK3 activity thus preventing the mitotic phosphorylation of CHMP4C.
The binding of HNF-1 to the putative promoter elements in mIMCD3 cells was confirmed by electrophoretic mobility shift assay (EMSA) (Figure 1a).
In addition, we found that all mutant constructs maintained their ability to bind DNA, except the N178Q DNA binding domain mutant which showed significantly reduced DNA binding as demonstrated by electrophoretic mobility shift assays (EMSA; Figure S1B; ref. 18).
To this end, we performed electrophoresis mobility shift assays (EMSAs; Figure 3) using 33P-end-labeled probes and whole cell extract prepared from cells over-expressing ectopic HA-MondoA and Myc-MLX.
As suggested by the reviewer, we have performed additional experiments to test the previously reported motif in an electrophoretic mobility shift assay (new Figure 7 figure supplement 1).
Fluorescently-labeled oligonucleotide probes representing each of these sites bound sequence-specifically to recombinant SpRunt-1 in an electrophoretic mobility shift assay (EMSA; Figure 4B).
Because of the nuclear dominant-negative activity we examined DNA-binding of the STAT3-YF-YFP-NLS mutant in stably transfected, inducible HEK cells by electrophoretic mobility shift assay (EMSA, Figure 5B).
A radioactively labelled 24 nt RNA probe spanning the putative hnRNPH binding site, incubated with nuclear extract, formed a slow-migrating complex, identified by electrophoretic mobility shift assay (EMSA; Figure 3D, lane 2).
