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. 2013 Jun 14;8(6):e68065. doi: 10.1371/journal.pone.0068065

Figure 4. FKHR-PAX3 localized predominantly in the cytoplasm of cells.

Figure 4

(A) NIH3T3 and RH30 cells were transfected with vectors expressing wild-type FKHR-GFP, triple-mutant (T24A/S256A/S319A) FKHR-GFP, or FKHR-PAX3-GFP (FP3-GFP) by lipofection. Cells were maintained in low serum medium (0.5% FBS) overnight prior to the addition of DMSO or LepB (2 μM) for six hours. At the end of treatment, fluorescent microscopy (magnification: 200X) was used to visualize the GFP-tagged proteins as indicated. (B) Western analysis confirmed the cytoplasmic localization of the FKHR-PAX3 protein in cells. RH30 cells were transfected with FKHR-PAX3 and treated with or without LepB as described in (4A). MyoG and α-tubulin served as nuclear and cytoplasmic specific controls, respectively, to evaluate the fractionation protocol. L27 antibody was used to detect FKHR-PAX3. Representative data from FKHR-PAX3 isoform c is shown.