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Figure 3 | Journal of Molecular Signaling

Figure 3

From: Estradiol effects on the dopamine transporter – protein levels, subcellular location, and function

Figure 3

PC12 cells have both membrane and intracellular ERα that is increased by NGFβ treatment. ERα was detected with C542 Ab. A. Protein levels of ERα in whole-cell extracts were determined by immunoblot analysis at 2, 4 and 7 days ± NGFβ treatment in medium lacking serum. Representative of 3 experiments. B. Micrograph of immunocytochemical staining of ERα using nonpermeabilized cells that had been serum-starved for 48 hours while being treated with 20 ng/ml NGFβ. The bar represents 2 μm. Fluorescent images viewed with an FITC filter were photographed. Left panel: Transmission micrograph of middle panel. Middle panel: Immunocytochemistry of ERα shown in red (Vector Red product), while the autofluorescent background is green. Right panel: Punctate ERα staining is irregularly distributed on the cell surface of a more highly magnified cell. C. The fixed cell microplate immunoassay shows that ERα is present in PC12 cells grown in serum-containing medium. The values for nonpermeabilized cells are shown by symbols: different ERα Ab concentrations, combined control conditions (IgG isotype control, no 1°, and no 1°/no 2°), superimposed at the origin as their values are all very low and do not differ from each other significantly, clathrin. The values for permeabilized cells are shown by bars at the appropriate Ab concentrations. The crosshatched bar is ERα, the gray bar at the origin represents combined controls (see above), and the white bar is clathrin (the permeabilization indicator). This graph represents average values from 3 experiments ± S.E.M.

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