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. 2017 Mar 30;45(11):6729–6745. doi: 10.1093/nar/gkx213

Figure 5.

Figure 5.

Decreased association of a Prp3 SUMOylation-deficient mutant with U2 and U5-containing RNP complexes. Overexpressed wt or 2KR T7-Prp3 was immunoprecipitated from HEK 293T cell lysates. (A) Co-IP of endogenous U2-SF3a120, U5-Snu114 and U4/U6-60K was assessed by western blot with antibodies specific for each protein and visualized with an Odyssey imaging system. (B) Quantification corresponding to three independent co-IP experiments as the one shown in (A) was performed with Image Studio Software (LI-COR Biosciences), according to the following calculation: ‘fold change’ = [IP/input]wt / [IP/input]2KR. (C) Co-IP of U4, U6, U2 or U5 snRNA was quantified by RT-qPCR with primers specific for each snRNA. Data are represented as mean ± S.E. (n = 3, ***P ≤ 0.001; Student's t test).