Figure 8.
G3BP1 and G3BP2 selectively associate with dissociated 40S ribosomal subunits. (A) Western blot of IPs from cells expressing either GFP (lanes 1–3) or RPS6-GFP (lanes 4–6), lysed in either ribosome-dissociating (EE) or ribosome-stabilizing (+5.0 mM MgCl2) buffer, washed, and incubated with RNase A or buffer. Released material was concentrated using acetone precipitation before SDS treatment; the bound material (GFP-IP) was eluted in SDS. IPs, lanes 1–6; released material, lanes 8–13. Mr (kD) are shown. (B) Stably expressed GFP-RPS6 in U2OS-WT cells, or GFP, GFP-tagged G3BP1 variants, or G3BP2a stably expressed in ΔΔG3BP1/2 cells were lysed in ribosome-dissociating EE or in ribosome stabilizing (EE + 7 mM MgCl2) buffer and immunoprecipitated and Western blotted as indicated. Mr (kD) are shown.

G3BP1 and G3BP2 selectively associate with dissociated 40S ribosomal subunits. (A) Western blot of IPs from cells expressing either GFP (lanes 1–3) or RPS6-GFP (lanes 4–6), lysed in either ribosome-dissociating (EE) or ribosome-stabilizing (+5.0 mM MgCl2) buffer, washed, and incubated with RNase A or buffer. Released material was concentrated using acetone precipitation before SDS treatment; the bound material (GFP-IP) was eluted in SDS. IPs, lanes 1–6; released material, lanes 8–13. Mr (kD) are shown. (B) Stably expressed GFP-RPS6 in U2OS-WT cells, or GFP, GFP-tagged G3BP1 variants, or G3BP2a stably expressed in ΔΔG3BP1/2 cells were lysed in ribosome-dissociating EE or in ribosome stabilizing (EE + 7 mM MgCl2) buffer and immunoprecipitated and Western blotted as indicated. Mr (kD) are shown.

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