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Figure 7. Shisa Inhibits Protein Maturation of FGFR (A) Shisa physically interacted with FGFR but not with ActRI or ActRII. Immunoprecipitation analysis was carried out as described in Figure 5. Cells were transfected in a 6-well plate with DNAs: fgfr-HA, 300 ng; ActRI-HA, 100 ng; ActRII-HA, 10 ng; shisa-Flag, 200 ng. (B) Coimmunoprecipitated FGFR-HA with Shisa was low molecular weight form only. (C–D‴) Confocal images of live HEK 293T cells show the cellular localization of FGFR-GFP (green) and DsRed-ER (Red). Cells were transfected in an 8-chamber slide with DNAs: fgfr-GFP, 20 ng; shisa, 50 ng; pDsRedER, 10 ng. (C–C‴) Transfectant of fgfr-GFP (cell surface expression of FGFR, n = 100, 95%). (D–D‴) Transfected fgfr-GFP with shisa (ER retention of FGFR, n = 100, 92%). (E) Shisa inhibited tyrosine-phosphorylation of FGFR. After stimulation with FGF2, lysate of cells expressing DNAs indicated at top was immunoprecipitated with rabbit α-HA Ab and was blotted with rat α-HA mAb (left) or mouse α-phosphotyrosine mAb (right). Image published in: Yamamoto A et al. (2005) Copyright © 2005. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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