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. 2020 Dec 24;49(2):1094–1113. doi: 10.1093/nar/gkaa1226

Figure 1.

Figure 1.

Identification of early assembly intermediates with NOP58 mutant proteins. (A) Epifluorescence microscopy images of HeLa cells expressing wild type (WT) or mutant GFP fusions of NOP58 (NOP58-KA/AR or NOP58-A283P). Blue/left panels: DAPI staining; Green/middle panels: GFP. Scale bar is 10 μm. (B) Schematic representation of SILAC IP experiments shown in C, D and E. First experiment was done with three conditions (L1: light label for control done with parental HeLa cells; M1: medium label for GFP-NOP58-WT; H1: heavy label for GFP-NOP58-A283P) and second with two conditions (L2: light label for control; M2: medium label for GFP-NOP58-KA/AR). (C–E) Proteomic analyses of the partners of GFP-NOP58-WT (C), GFP-NOP58- A283P (D) and GFP-NOP58-KA/AR (E). Graphs display SILAC ratios (y axis, specific versus control IP) as a function of signal abundance (x axis, log10(intensity)). Each dot represents a protein and is color encoded according to the classification shown between panels C and D. The labeled dots highlight proteins relevant to this study. Full hit list with Significance B values are given in Supplementary Table S1. (F) Bar plot comparing SILAC ratios shown in C, D and E for a few selected partners. In each experiment, the normalization is done by dividing the SILAC ratio of the protein of interest with that of NOP58. Blue bars are for NOP58-A283P, red bars for NOP58-KA/AR and green bars for NOP58-WT.