Fig. 5.

SUMOylation does not alter ING1b localization. (A) U2OS cells transfected with control vector or either ING1b- or SUMO-deficient mutants (K193R or E195A) were stained with αING1 or DAPI to stain ING1b protein and DNA, respectively. Cells expressing ING1b, ING1b K193R and ING1b E195A were imaged with the same exposure time and for cells expressing control vector, exposure time was increased in order to detect endogenous ING1b. (B) The chromatin-enriched fractions (CEFs) or whole cell extracts (WCE) from U2OS cells transfected with empty vector, ING1b WT or SUMO-specific mutant (ING1b E195A) were subjected to IB using αING1 to detect chromatin bound ING1b and αH3 for establishing equal loading of CEFs. WCE was probed for αING1 and with actin to confirm equal protein expression and loading, respectively.