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. 2013 Feb 7;8(2):e55342. doi: 10.1371/journal.pone.0055342

Figure 4. Localization of GFP-dFMRP fusion proteins in SG as analyzed in fixed cells.

Figure 4

(A) Schematic representation of GFP-dFMRP (top) and its deletion versions. (B) Schneider cells were transfected with either GFP or GFP-dFMRP constructs for 48 h. Cells-expressing GFP-dFMRP were then collected and protein extracts were next analyzed by immunoblotting for GFP-dFMRP expression using anti-GFP antibodies. Tubulin was used as a loading control. (C–D) Schneider cells were transfected with either GFP or GFP-dFMRP constructs for 48 h. Cells were then fixed and then processed for immunofluorescence to detect GFP or GFP-dFMRP (green). The intracellular localization of endogenous dPABP (C; red) is revealed using antibodies specific to dPABP. The indicated percentage of dFMRP-granules harboring dPABP (D) is calculated from 3 different experiments containing a total of 500 transfected cells. Scale bars are indicated. (E) Schneider cells were transfected with either GFP or GFP-dFMRP constructs for 48 h. Cells were then treated with arsenite (0.5 mM; 1.5 h), fixed and processed for immunofluorescence to detect dPABP using specific antibodies (red signal). GFP and GFP-dFMRP are detected as green fluorescence. Scale bars are indicated.