Figure 4. Ssd1 coprecipitates with P-body and stress granule proteins. Ssd1-GFP was immunoprecipitated from extracts of logarithmically growing yeast expressing TAP-tagged P-body or stress granule proteins and Ssd1-GFP. Immunoblots were probed with anti-GFP or anti-TAP antibodies. Input = immunoblots of total cell extract. Lanes 2, 4, 6, and 8 are from cells expressing TAP-tagged proteins only (Dcp2, Not1, Xrn1, Pat1). Lanes 1, 3, 5, 7, and 9 also contain pGPD-Ssd1-GFP (FLE1019). Dcp2, Not1, Xrn1, and Pat1 coprecipitate with Ssd1-GFP. Note that a trace of Not1-TAP nonspecifically precipitates in the absence of Ssd1-GFP (lane 4); however, much more coprecipitates with Ssd1-GFP. Control experiments (not depicted) confirm that Ssd1 coprecipitates with Cbk1, as anticipated by previously published two-hybrid assays (Racki et al., 2000).
Figure 4.

Ssd1 coprecipitates with P-body and stress granule proteins. Ssd1-GFP was immunoprecipitated from extracts of logarithmically growing yeast expressing TAP-tagged P-body or stress granule proteins and Ssd1-GFP. Immunoblots were probed with anti-GFP or anti-TAP antibodies. Input = immunoblots of total cell extract. Lanes 2, 4, 6, and 8 are from cells expressing TAP-tagged proteins only (Dcp2, Not1, Xrn1, Pat1). Lanes 1, 3, 5, 7, and 9 also contain pGPD-Ssd1-GFP (FLE1019). Dcp2, Not1, Xrn1, and Pat1 coprecipitate with Ssd1-GFP. Note that a trace of Not1-TAP nonspecifically precipitates in the absence of Ssd1-GFP (lane 4); however, much more coprecipitates with Ssd1-GFP. Control experiments (not depicted) confirm that Ssd1 coprecipitates with Cbk1, as anticipated by previously published two-hybrid assays (Racki et al., 2000).

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