Additional file 8 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 7: Fig. S6. Effect of NOLC1 and TCOF1 knock-down on the nucleolar localization of FGF12 and its interaction with NOLC1 and TCOF1. HEK 293 cells were transfected with NOLC1/TCOF1-targeting siRNA or scramble siRNA (control). Fluorescence images of in situ PLA using a mixture of rabbit anti-FGF12 and mouse anti-NOLC1 or anti-TCOF1 antibodies. Cell nuclei were labeled with NucBlue Live and cells were analyzed with fluorescence microscopy. The dashed line indicates the cell area. The scale bar represents 20 μm

Additional file 7 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 6: Fig. S5. Efficiency of NOLC1, TCOF1 and FGF12 knock-down in U2OS-FGF12-mGFP-myc cells. Western blotting analysis of cell lysates of U2OS-FGF12-mGFP-myc cells treated with siRNA against NOLC1, TCOF1 or FGF12

Additional file 4 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 3: Fig. S2. Streptavidin agarose pull-down from U2OS cells stably expressing FGF12-SBP. A Western blotting of cell lysates of U2OS-FGF12-SBP and U2OS cells (control) to confirm FGF12-SBP expression. B U2OS-FGF12-SBP and U2OS cells were lysed and co-purification of NOLC1 and TCOF1 with FGF12-SBP on streptavidin agarose was verified by SDS-PAGE and western blotting. C Streptavidin-agarose pull-down from U2OS-FGF12-SBP cells to purify FGF12-SBP complexes under native conditions. Bound proteins were eluted with biotin and separated by 2D-BN-PAGE. Co-migration of FGF12-SBP and NOLC1 or TCOF1 was determined by western blotting

Additional file 2 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 1: Table S1. Results of MS-based peptide identification for proteins that bind specifically to FGF12-His in pull down experiments. ID is the Uniprot protein identifier. The score and peptide match for each of two repeats of the experiment are given

Additional file 3 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 2: Fig. S1. Figure S1. Cellular localization of FHF proteins. A Western blotting analysis of whole cell lysates detecting mGFP in U2OS cells stably transfected with FGF11-mGFP-myc, FGF12-mGFP-myc, FGF13-mGFP-myc, FGF14-mGFP-myc or U2OS cells (control) to confirm the expression of fusion proteins. B Localization of FHF proteins in U2OS cells stably transfected with FGF11-mGFP-myc, FGF12-mGFP-myc, FGF13-mGFP-myc and FGF14-mGFP-myc. Nuclei were labeled with NucBlue Live and cells were analyzed with fluorescence microscopy. The dashed line indicates the cell area. The scale bar represents 20 μm. C Quantification of the amount of fluorescent proteins in each compartment (cytoplasm, nucleus, nucleolus) including mean fluorescence intensity and compartment area. Data presented are means ± SD of 20 cells. Student's t-test was used for statistical analysis; ***p < 0.001; **p < 0.01. D U2OS-FGF11-mGFP-myc, U2OS-FGF12-mGFP-myc, U2OS-FGF13-mGFP-myc and U2OS-FGF14-mGFP-myc cells were washed, lysed and fractionated into cytoplasmic and nuclear fractions. FHFs-mGFP-myc were extracted from each fraction by adsorption onto anti-myc resin, separated by SDS-PAGE and analyzed by western blotting. ERK1/2 and histone H3 served as cytoplasmic and nuclear marker proteins, respectively

Additional file 5 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 4: Fig. S3. Negative controls for in situ PLA performed in U2OS-FGF12-mGFP-myc cells. PLA in U2OS-FGF12-mGFP-myc cells treated with a single antibody or in untreated cells. Nuclei were labeled with NucBlue Live and cells were analyzed with fluorescence microscopy. Scale bar represents 20 μm

Additional file 6 of FGF12 is a novel component of the nucleolar NOLC1/TCOF1 ribosome biogenesis complex

Additional file 5: Fig. S4. PLA in HEK 293 cells lacking ectopic FGF12. Fluorescence images of in situ PLA using rabbit anti-FGF12 and mouse anti-NOLC1 or anti-TCOF1 antibody in HEK 293 cells. HEK 293 cells treated with a single antibody or untreated showed no signal and served as negative controls. Nuclei were labeled with NucBlue Live and cells were analyzed with fluorescence microscopy. The dashed line indicates the cell area. The scale bar represents 20 μm