Mass Spectroscopy analyses for novel RBPs: a Protein elute from RaPID was subjected to PAGE electrophoresis and silver-stained. The areas marked by squares were excised from the gel and subjected to mass spectroscopy. Protein size markers (in kDa) are indicated to the left. Open arrow indicates the MS2 fusion protein. b The proteins with significant signals in the mass spec. analysis (Additional file 1: Table S1) were grouped according to their function. c RaPID was performed to cells with either untagged PMP1 (−MS2L) or tagged mRNA (+MS2L). Protein samples were resolved on PAGE, silver stained, and bands that appeared stronger in the +MS2L (marked by squares) were excised and subjected to mass spec. Protein size markers (in kDa) are indicated to the left.
Open arrow indicate the band corresponding in size to MS2 fusion protein. d Western analysis confirmation for Yef3 association with PMP1. RaPID was performed to the indicated strains and samples from the Input and the Elution were subjected to western analysis with the indicated antibodies. Note that Yef3 is usually detected as two bands due to cleavage