Figure 7

Hyaluronan/RNA-binding assays. To confirm the interaction of USP17 subfamily proteins with either hyaluronan or RNA, CPC precipitation assay was performed. Cell lysate aliquots from HeLa cells transfected with myc-tagged USP17L or myc-tagged USP17N construct were incubated with hyaluronan or RNA and subjected to CPC precipitation. (A) Coprecipitated proteins were subjected to Western blot analysis with an antibody against the myc tag. CPC precipitation without the addition of CPC or myc-tagged USP17 construct was used as negative controls (lanes 1–3). Without the addition of CPC, the whole cell lysate from cells transfected with myc-tagged USP17L or myc-tagged USP17N was used as a positive control (lane 7). (B) For RNA binding assay, total RNA of HeLa cells instead of hyaluronan were added.