Fig. 3: RNase and ionic strength susceptibilities of Exon Junction Complexes.

a, b Lysates from cells coexpressing MAGOH-LgBiT and eIF4A3-SmBiT (OB9 cells) are fractionated on sucrose gradients. Luciferase activity (RLUs) and SYBR green fluorescence at 540 nm are measured in gradient fractions. a Control lysates (black), lysates with NaCl (600 mM) (light blue), b RNase (purple) or both RNAse and NaCl (orange) are added to lysates from EJC-NanoBiT expressing cells. c Luciferase activity in crude lysates supplemented with 600 mM NaCl (blue) or NaCl and RNase (orange). RNA in 3a, 3b and 3c are detected by SYBR green fluorescence emission at 540 nm (dots) and NanoBiT luciferase activities (continuous lines) are measured in 48 × 0.25 ml gradient fractions. d Lysates from HEK293 cells are fractionated on 11-54 % sucrose gradients. EJC core proteins are detected by Western blot in input fractions (top) or co-immunoprecipitated from the same fractions by anti-eIF4A3 (below). eIF4A3, MAGOH and Y14 proteins are detected on Western blots with specific antibodies diluted 1/1000. All these experiments have been repeated twice with similar results. Source data are provided in the Source Data file.