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. 2016 Apr 25;113(19):5447–5452. doi: 10.1073/pnas.1522458113

Fig. 4.

Fig. 4.

The recruitment of Prp19C/NTC to the spliceosome is impaired in atprmt5 mutants and restored in atprmt5-2 prp8-8. (A) The association of Prp19C/NTC and Prp8 (and Brr2) decreased in atprmt5 mutants and was rescued in atprmt5-2 prp8-8 mutants. Nuclear extracts from Col, atprmt5-2, atprmt5-2 prp8-8, and prp8-8 were immunoprecipitated by anti-Prp8 (Upper) and anti-Brr2 (Lower) polyclonal antibodies, and the interacting proteins were identified by MS. The spectral counts of Prp19C/NTC from atprmt5-2, atprmt5-2 prp8-8, and prp8-8 were divided by that from Col, and the ratios are shown in the histogram. (B and C) Prp8- or Brr2-associated proteins were identified in Col, atprmt5, atprmt5-2 prp8-8, and prp8-8 by MS. Prp8 (B) and Brr2 (C) were immunoprecipitated by anti-Prp8 and anti-Brr2 antibodies, respectively, and immunoblotting was performed with anti-MAC3A and anti-MOS4 antibodies for Prp19C/NTC. The dividing lines in C, Right indicate noncontiguous images between Col and prp8-8.