Effects of KREPB6, KREPB7, or KREPB8 repression on tagged KREN3, KREN2, or KREN1 editosomes, respectively. (A) Western analysis using an MAb mixture (upper panel) and SYPRO-Ruby stained gels (lower panel) of editosomes that were purified via the TAP-tag from total cell lysates. Editosomes were purified by sequential IgG Sepharose and calmodulin affinity chromatography from equal numbers of KREPB6 RNAi-N3TAP, KREPB7 RNAi-N2TAP, KREPB8 RNAi-N1TAP cells in which the KREPB protein expression was repressed by RNAi for 6 d (+) or were nonrepressed (−). (B–D) Western analysis of the glycerol gradient fractions of the complexes purified via IgG affinity chromatography and TEV protease cleavage from these cells.
