Figure 7.

Interaction-deficient OTUD5 mutants cannot rescue the transcription repressive phenotypes. The PTuner 263 cells expressing indicated OTUD5 constructs (EV = empty vector) were transfected with siRNAs targeting the 3’UTR region of OTUD5, treated with 4-OHT and Shield-1 to stabilize Fok1, then the presence of YFP-MS2 expression at the Fok1 spots was measured. YFP-MS2 reporter was induced by tetracycline 3 hours prior to fixing. Only the cells with positive FLAG spots were counted. The assay was performed in triplicates (N = 50). On the center is the quantification of the MS2 RFI at Fok1 nuclease using Image J (N = 50). Below is the expression confirmation of each cell. Scale bars indicate 10 μm. (**** indicates P-value < 0.0005, ** <0.05.)