G3BP1 RGG motif is required for association with 40S ribosomal subunits and for SG competence. (A) GFP and GFP-tagged G3BP1 variants were expressed in COS7 cells, lysed in EE buffer, and immunoprecipitated with RNase treatment. Lysates and IPs were resolved using SDS-PAGE and subjected to Western blotting for the indicated proteins. Mr (kD) are shown. (B) GFP and GFP-tagged G3BP1 variants were stably expressed in ΔΔG3BP1/2 cells and immunoprecipitated. Lysates and IPs were resolved using SDS-PAGE and subjected to Western blotting for the indicated proteins. Mr (kD) are shown. (C) ΔΔG3BP1/2 cells expressing the indicated proteins were treated as indicated, stained, and scored for SGs using eIF4G and eIF3b. Data shown are mean ± SEM and are analyzed using the unpaired t test, WT versus G3BP1 variants. ***, P < 0.005; n = 3.
Figure 7.

G3BP1 RGG motif is required for association with 40S ribosomal subunits and for SG competence. (A) GFP and GFP-tagged G3BP1 variants were expressed in COS7 cells, lysed in EE buffer, and immunoprecipitated with RNase treatment. Lysates and IPs were resolved using SDS-PAGE and subjected to Western blotting for the indicated proteins. Mr (kD) are shown. (B) GFP and GFP-tagged G3BP1 variants were stably expressed in ΔΔG3BP1/2 cells and immunoprecipitated. Lysates and IPs were resolved using SDS-PAGE and subjected to Western blotting for the indicated proteins. Mr (kD) are shown. (C) ΔΔG3BP1/2 cells expressing the indicated proteins were treated as indicated, stained, and scored for SGs using eIF4G and eIF3b. Data shown are mean ± SEM and are analyzed using the unpaired t test, WT versus G3BP1 variants. ***, P < 0.005; n = 3.

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