Fig. 5

UCHL1 interacts with the shelterin complex and the nuclear scaffold. a DU 145 or HEK293T cellular lysates in a low stringency buffer were incubated with anti-TRF2 antibodies or control IgG. The immunoprecipitate (IP), and equal volumes of lysate (Input) and immunodepleted (ID) fractions were analyzed by immunoblotting with the indicated antibodies. b In left panel, DU 145 cellular lysate in a low stringency buffer was incubated with anti-p53 antibodies or control IgG. In right panel, cell lysate in a high stringency buffer of DU 145 cells treated with the cross-linker DSP was incubated with anti-UCHL1 antibodies or control IgG. The immunoblot analysis was done as in (a). c In situ PLA images of the interaction between TRF2 and RAP1 (positive control), UCHL1 and RAP1 or UCHL1 and TRF2 in HEK293T cells. PLA signals appear as discrete red dots and nuclei are visualized by DAPI (blue). A total of 30 nuclei per group were quantified. The average number of PLA foci per nucleus was graphed with error bars representing standard errors of the means. Single primary antibodies, isotype control and PLA probes only were used as negative controls as indicated. ****p < 0.0001 (determined by one-way ANOVA)