Fig. 1.
Setup of mRNA interactome capture. A) Detailed schematic of the interactome capture protocol. B) mRNA recovery of a poly-adenylated 32P-body-labeled beta-globin RNA. Scintillation counts in lysates or supernatants of each step of the protocol are shown. After addition of the spike into whole cell lysates, capture was performed +/− LiDS in lysis and wash buffer. C) Protein recovery of 35S-Methionine-labeled proteins. Scintillation counts of 35S levels of supernatants are shown for individual steps of the protocol. D) qRT-PCR of actin (ACT1), GAPDH (TDH3) and ribosomal RNA (18S rRNA) from cells lacking UV irradiation (-UV), 0.72 or 7.2 J/cm2 of 365 nm UV light exposure. Input was adjusted to 100% and recovered mRNA levels are shown as fraction of the input. Error bars in panels B, C and D represent standard deviation (SD), n = 3.